Open Access

Gender-specific effects of intrauterine growth restriction on the adipose tissue of adult rats: a proteomic approach

  • Adriana Pereira de Souza1,
  • Amanda Paula Pedroso1,
  • Regina Lúcia Harumi Watanabe1,
  • Ana Paula Segantine Dornellas1,
  • Valter Tadeu Boldarine1,
  • Helen Julie Laure2,
  • Claudia Maria Oller do Nascimento1,
  • Lila Missae Oyama1,
  • José Cesar Rosa2 and
  • Eliane Beraldi Ribeiro1Email author
Proteome Science201513:32

https://doi.org/10.1186/s12953-015-0088-z

Received: 7 August 2015

Accepted: 26 November 2015

Published: 2 December 2015

Abstract

Background

Intrauterine growth restriction (IUGR) may program metabolic alterations affecting physiological functions and lead to diseases in later life. The adipose tissue is an important organ influencing energy homeostasis. The present study was aimed at exploring the consequences of IUGR on the retroperitoneal adipose tissue of adult male and female rats, using a proteomic approach.

Methods and Results

Pregnant Wistar rats were fed with balanced chow, either ad libitum (control group) or restricted to 50 % of control intake (restricted group) during the whole gestation. The offspring were weaned to ad libitum chow and studied at 4 months of age. Retroperitoneal fat was analyzed by two-dimensional gel electrophoresis followed by mass spectrometry.

Both male and female restricted groups had low body weight at birth and at weaning but normal body weight at adulthood. The restricted males had normal fat pads weight and serum glucose levels, with a trend to hyperinsulinemia. The restricted females had increased fat pads weight with normal glucose and insulin levels.

The restricted males showed up-regulated levels of proteasome subunit α type 3, branched-chain-amino-acid aminotransferase, elongation 1- alpha 1, fatty acid synthase levels, cytosolic malate dehydrogenase and ATP synthase subunit alpha. These alterations point to increased proteolysis and lipogenesis rates and favoring of ATP generation.

The restricted females showed down-regulated levels of L-lactate dehydrogenase perilipin-1, mitochondrial branched-chain alpha-keto acid dehydrogenase E1, and transketolase. These findings suggest impairment of glycemic control, stimulation of lipolysis and inhibition of proteolysis, pentose phosphate pathway and lipogenesis rates.

In both genders, several proteins involved in oxidative stress and inflammation were affected, in a pattern compatible with impairment of these responses.

Conclusions

The proteomic analysis of adipose tissue showed that, although IUGR affected pathways of substrate and energy metabolism in both males and females, important gender differences were evident. While IUGR males displayed alterations pointing to a predisposition to later development of obesity, the alterations observed in IUGR females pointed to a metabolic status of established obesity, in agreement with their increased fat pads mass.

Keywords

Intrauterine growth restriction Maternal undernutition Adipose tissue Proteome

Background

The concept of fetal programming describes that the exposition to adverse stimuli or insults, during critical phases of intrauterine development, may induce permanent changes in physiological functions and lead to adulthood diseases [1]. Increased risks of type 2 diabetes, insulin resistance, cardiovascular diseases and obesity have been associated with intrauterine growth restriction (IUGR) induced by undernutrition [2, 3]. These consequences have been shown to depend on the severity, duration and gestational period of the insult and also to be gender-dependent [4, 5]. Importantly, a mismatch between intrauterine and post-natal nutritional environment has been shown to be relevant for the expression of the programmed metabolic dysfunctions [6].

Previous reports have found that the adult offspring of IUGR rats displayed hyperphagia, obesity, hypertension, high serum leptin and insulin levels, increased hypothalamic density of leptin and serotonin receptors, and impairment of serotonin and insulin hypothalamic signaling [713]. Additionally, IUGR led to increased circulating levels of catecholamines in rats [14] and decreased levels of branched-chain amino acids in mice [15].

Lipogenesis and lipolysis are important physiologic pathways in the adipose tissue. Fatty acids for triacylglycerols synthesis may be taken up from the circulation or derive from de novo synthesis from glucose. Glucose degradation also yields glycerol 3-phosphate for fatty acids esterification and storage [16, 17]. Conversely, fatty acids and glycerol derived from triacylglycerols lipolysis may be released into the circulation. Those are hormone-controlled pathways. Lipolysis is inhibited by insulin and stimulated by cathecolamines and growth hormone. On the other hand, lipogenesis is stimulated by insulin while growth hormone is inhibitory [1618]. Rat studies have indicated that IUGR due to maternal food restriction decreased adipose tissue lipolysis while it increased lipogenesis and/or adipogenesis, due to impairment of sympathetic activity [2].

The adipose tissue is also an endocrine organ whose secretions influence the onset of metabolic disorders [19]. Increased production of pro-inflammatory adipokynes in obesity plays a relevant role in the linking of adiposity, metabolic syndrome and cardiovascular diseases [20, 21]. Recent reports have shown that fetal leptin and adiponectin levels closely related to birth weight and IUGR has been shown to increase leptin but not adiponectin levels. Moreover, TNF-α levels have been found to be either normal or increased while IL-6 levels were either increased or decreases in IUGR [19, 22, 23].

Proteomic analysis allows the exam of hundreds of proteins in a sample and the identification of modification on their expression pattern in response to physiologic, pathologic and nutritional alterations, possibly leading to the identification and characterization of biological markers [24, 25]. Two-dimensional gel electrophoresis (2DE) followed by mass spectrometry remains an effective methodology in proteomics, especially as an initial approach [26, 27]. Recent proteomic studies have focused on the consequences of IUGR in tissues of animals and humans. Down-regulation of proteins related to oxidative phosphorylation has been found in the liver of both male and female IUGR rats [28]. In piglets, IUGR up-regulated subcutaneous adipose tissue levels of proteins related to glucose and fatty acid metabolism, lipid transport and apoptosis [29]. In humans, IUGR has been shown to increase serum levels of proteins related to signal transduction, blood coagulation and antioxidant response, while immune response proteins were down-regulated [30].

The above data indicate that IUGR may injure multiple aspects pertinent to adipocytes physiology that are relevant to the development of metabolic impairment and obesity in adulthood. Considering the above, the objective of this study was to further explore the consequences of IUGR in the adipose tissue of adult rats through proteomic approach.

Results

Body and white adipose tissue weight and blood and tissue parameters

Restricted male and female rats had low body weight at birth and at weaning but this difference was no longer observed at four months of age (Table 1). Food intake was similar between control and restricted animals, from weaning to 4 months of age (data not shown).
Table 1

Body and white adipose tissue weight of male and female control and restricted offspring

 

Male

Female

Control (16)

Restricted (17)

Control (14)

Restricted (14)

BW at birth (g)

6.08 ± 0.11

4.84 ± 0.12***

5.69 ± 0.14

4.80 ± 0.13***

BW at weaning (g)

85.44 ± 2.32

76.05 ± 2.52**

79.83 ± 2.15

70.24 ± 2.68**

BW at 4-months (g)

390.5 ± 7.2

385.4 ± 10.6

234.9 ± 3.3

227.2 ± 4.2

BW gain (g)

263.6 ± 8.4

271.2 ± 10.8

121.2 ± 4.8

127.2 ± 5.5

Retroperitoneal (g/100 g bw)

1.15 ± 0.11

1.24 ± 0.11

0.80 ± 0.06

0.95 ± 0.06

Mesenteric (g/100 g bw)

0.91 ± 0.07

0.98 ± 0.07

1.01 ± 0.05

1.19 ± 0.04**

Gonadal

1.31 ± 0.11

1.50 ± 0.08

2.46 ± 0.19

3.0 ± 0.17*

Total weight (g/100 g bw)

3.37 ± 0.28

3.72 ± 0.23

4.27 ± 0.28

5.22 ± 0.22*

Data are means ± SEM; (number of animals)

BW body weight, g/100 g bw grams/100 g of body weight, Total weight sum of retroperitoneal, mesenteric and gonaldal fat pads

*p < 0.01 vs. control

**p < 0.01 vs. control

***p < 0.001 vs. control

White adipose tissues weight were similar between control and restricted males. The female restricted rats showed increased weight of mesenteric and gonadal white adipose tissue and the sum of the three fat pads was higher than that of the control females (Table 1).

Serum glucose, adiponectin, corticosterone and triglycerides were similar between the groups of male rats. Serum insulin levels showed a tendency to increase in the restricted males (p = 0.083). Serum and tissue cytokines levels were similar between the male groups (Table 2).
Table 2

Blood and retroperitoneal adipose tissue parameters of adult male and female control and restricted offspring

 

Male

Female

Control

Restricted

Control

Restricted

Serum glucose (mg/dl)

100.2 ± 5.2 (18)

92.4 ± 3.1 (9)

95.2 ± 3.9 (13)

100.0 ± 5.2 (9)

Serum Insulin (ng/ml)

0.50 ± 0.07 (8)

0.79 ± 0.13 (9)

0.35 ± 0.06 (9)

0.34 ± 0.12 (9)

Serum adiponectin (μg/ml)

12.3 ± 1.4 (9)

11.3 ± 1.1 (9)

17.45 ± 1.81 (9)

18.47 ± 2.27 (9)

Serum corticosterone (ng/ml)

94.2 ± 7.9 (9)

97.0 ± 8.8 (8)

63.58 ± 9.54 (9)

71.06 ± 5.58 (9)

Serum triglycerides (mg/dl)

55.1 ± 4.3 (18)

68.6 ± 9.3 (9)

37.2 ± 3.1 (14)

38.7 ± 6.0 (8)

Serum TNF-α (pg/ml)

7.6 ± 1.7 (8)

18.2 ± 6.4 (8)

20.07 ± 1.8 (8)

14.78 ± 4.3 (9)

Serum IL-1β (ng/ml)

0.10 ± 0.04 (6)

0.06 ± 0.01 (6)

0.14 ± 0.04 (9)

0.04 ± 0.01*(8)

Tissue TNF- α (pg/ml)

49.3 ± 4.5 (8)

55.0 ± 3.2 (8)

63.93 ± 4.9 (8)

71.55 ± 4.3 (8)

Tissue IL-6 (pg/ml)

107.3 ± 7.0 (8)

106.6 ± 6.9 (8)

126.24 ± 3.8 (8)

134.13 ± 6.9 (8)

Tissue IL-10 (pg/ml)

187.6 ± 19.2 (8)

225.0 ± 28.7 (8)

275.89 ± 19.5 (8)

311.09 ± 19.5 (8)

Data are means ± SEM. (number of animals)

*p < 0.05 vs control

For female rats, no differences were found in serum glucose, insulin, corticosterone, adiponectin and triglycerides between the control and restricted groups. The restricted females showed low serum IL-1β (Table 2).

Proteomic analysis

The 2DE gels of retroperitoneal adipose tissue showed 425 ± 2.9 spots in control males (N = 8) and 417 ± 4.5 spots in restricted males (N = 8). Of these, 37 spots showed significant density changes, with 15 spots under- and 22 over-expressed. Spots optic densities are shown in Additional file 1: Table S1. The significantly affected spots were analyzed by mass spectrometry for proteins identification. Figure 1 shows a representative image of a 2DE gel of a control male with indication of the spots significantly affected by IUGR.
Fig. 1

Representative image of a 2DE gel of a control male retroperitoneal adipose tissue depicting the proteins significantly affected by IUGR. The numbers indicate the protein acession number. Numbers in squares indicate the over-expressed identified proteins. Numbers in circles indicate the under-expressed identified proteins

The MS analysis identified 11 of the 15 under-expressed proteins and 17 of the 22 over-expressed proteins. One down-regulated protein (Murinoglobulin-1) and 2 up-regulated proteins (Actin, cytoplasmic I and Voltage-dependent anion-selective channel protein 1) were identified in 2 adjacent spots. Table 3 shows the Swiss-Prot Accession Numbers (available at http://www.expasy.ch/sprot), full protein names, theoretical molecular weight (MW) and isoelectric point as well as the mass spectrometry data of the identified proteins having statistically significant Mascot score results (p < 0.05) in males. Additional file 2: Table S2 shows gene names and biological processes of the proteins significantly up-regulated and down-regulated proteins, as assessed by Panther software. Metabolic process was the most common biological process class for both the down-regulated (7 out of 10) and the up-regulated (9 out of 15) proteins. The metabolic processes included lipidic, amino acid and carbohydrate metabolism (Table 4).
Table 3

Identified proteins with significant expression alteration between control and restricted males

Accession Number

Protein Name

Matched Peptides

Score

Coverage (%)

Fold Change (R/C)

MW (Da)/ pI

Down-regulated Proteins

 P06761

78 kDa glucose-regulated protein

6

334

11

0.60

71476/5.07

 P14668

Annexin A5

6

338

18

0.39

35780/4.93

 P34058

Heat shock protein HSP 90 β

9

245

12

0.68

83577/4.97

 P20059

Hemopexin

4

66

6

0.55

52072/7.58

 Q6AYC4

Macrophage-capping protein

1

50

3

0.63

39065/6.11

 Q03626

Murinoglobulin-1

3

66

3

0.47

166614/5.68

 Q03626

Murinoglobulin-1

2

81

1

0.24

166614/5.68

 Q63598

Plastin-3

1

46

1

0.22

71157/5.32

 P67779

Prohibitin

4

196

17

0.58

28860/5.57

 P09006

Serine protease inhibitor A3N

3

109

6

0.37

46796/5.33

 P48721

Stress-70 protein, mitochondrial

3

66

6

0.52

74102/5.97

Up-regulated Proteins

 P60711

Actin, cytoplamic I

6

191

21

1.77

42058/5.29

 P60711

Actin, cytoplamic I

4

244

15

1.83

42058/5.29

 P39069

Adenylate kinase izoenzyme 1

2

30

12

1.96

21686/7.66

 P07943

Aldose reductase

3

94

10

2.05

36238/6.26

 P15999

ATP synthase subunit alpha, mitochondrial

1

57

1

1.94

59833/9.22

 O35854

Branched-chain-amino-acid aminotransferase. mitochondrial

4

52

9

1.67

44827/8.46

 P62630

Elongation factor 1-alpha 1

2

52

4

1.79

50430/9.10

 P85845

Fascin

3

39

7

2.27

55211/5.96

 P12785

Fatty acid synthase

5

151

2

2.40

275146/5.96

 P05065

Fructose-biphosphate aldolase A

7

245

17

1.49

39791/8.31

 P20761

Ig gamma-2B chain C region

2

38

4

2.07

37112/7.70

 O88989

Malate dehydrogenase, cytoplasmic

3

81

11

2.44

36634/6.16

 P18422

Proteasome subunit α-type 3

4

86

15

1.93

28633/5.29

 P27867

Sorbitol dehydrogenase

2

94

7

2.05

38790/7.14

 P68370

Tubulin α-1A chain

3

121

9

2.37

50800/4.94

 Q9Z2L0

Voltage-dependent anion-selective channel protein 1

2

60

8

2.16

30853/8.62

 Q9Z2L0

Voltage-dependent anion-selective channel protein 1

1

42

4

4.94

30853/8.62

Accession number, protein name, number of matched peptides, proein score, percentage coverage, fold change (restricted/control) and theoretical molecular mass (Da) and pI of identified proteins

Table 4

Biological process classification of identified proteins of male rats

Protein name

Metabolic process

Down-regulated in restricted males

 Serine protease inhibitor A3N

Proteolysis

 Murinoglubulin-1

 Hemopexin

 78 kDa glucose-regulated protein

Protein folding

 Heat shock protein HSP 90-beta

 Prohibitin

DNA replication

 Annexin A5

Lipidic

Up-regulated in restricted males

 Branched-chain-amino-acid aminotransferase, mitochondrial

Amino acid

 Proteasome subunit α type-3

Proteolysis

 Fatty acid synthase

Lipidic

 Sorbitol dehydrogenase

Carbohydrate

 Malate dehydrogenase, cytoplasmic

Carbohydrate, tricarboxilic acid cycle (TCA)

 ATP synthase subunit alpha, mitochondrial

Respiratory chain

 Adenylate kinase isoenzyme 1

Nucleotide

 Elongation factor 1-alpha 1

Translation

 Aldose reductase

Transport

The 2DE gels of females had 404 ± 3.6 spots in the controls (N = 6) and 397 ± 3.9 spots in the restricted ones (N = 6). Of these, 27 spots showed significant density changes, with 20 spots under- and 7 over-expressed. Spots optic densities are shown in Additional file 1: Table S1. The significantly affected spots were analyzed by mass spectrometry for proteins identification. Figure 2 shows a representative image of a 2DE gel of a control female with indication of the spots significantly affected by IUGR.
Fig. 2

Representative image of a 2DE gel of a control female retroperitoneal adipose tissue depicting the proteins significantly affected by IUGR. The numbers indicate the protein acession number. Numbers in squares indicate the over-expressed identified proteins. Numbers in circles indicate the under-expressed identified proteins

The MS analysis identified 18 of the 20 down-regulated proteins and 4 of the 7 up-regulated proteins. Two down-regulated proteins (Serotransferrin and Ig gamma 2-A chain C) were identified in 2 adjacent spots. Female proteins data of significant Mascot score results (p < 0.05) are shown in Table 5. Additional file 3: Table S3 shows gene names and biological processes of the significantly up-regulated and down-regulated proteins in females. Metabolic process was the most common biological process class for the down-regulated (13 out of 16) and the up-regulated (4 out of 4) proteins. The metabolic processes included lipidic, amino acid and carbohydrate metabolism (Table 6).
Table 5

Identified proteins with significant expression alteration between control and restricted females

Accession number

Protein name

Matched peptides

Score

Coverage (%)

Fold change (R/C)

MW (Da)/ pI

Down-regulated proteins

 F1LMZ8

26S proteasome non-ATPase regulatory subunit 11

2

89

4

0.44

47724/6.08

 P35738

2-oxoisovalerate dehydrogenase subunit β. mitochondrial

1

45

4

0.46

43550/6.41

 P30713

Glutathione S-transferase theta-2

1

47

5

0.38

27596/7.75

 A7VJC2

Heterogeneous nuclear ribonucleoproteins A2/B1

4

96

13

0.35

37513/8.97

 P20760

Ig gamma 2-A chain C

3

79

9

0.53

35685/7.72

 

Ig gamma 2-A chain C

4

109

13

0.23

 

 P42123

L-lactate dehydrogenase B

1

30

2

0.57

36879/5.70

 P43884

Perilipin 1

2

74

5

0.41

55986/6.37

 Q9Z1H9

Protein kinase C delta binding protein

5

119

18

0.33

27894/5.79

 P62836

Ras-related protein Rap-1A

1

39

5

0.32

21322/6.38

 P05545

Serine protease inhibitor A3K

3

119

10

0.45

46764/5.31

 P05544

Serine protease inhibitor A3L

1

46

2

0.29

46442/5.48

 P12346

Serotransferrin

6

133

8

0.38

78550/7.14

 P12346

Serotransferrin

2

110

3

0.31

78550/7.14

 Q66X93

Staphylococcal nuclease domain-containing protein 1

3

38

4

0.39

103585/6.76

 P50137

Transketolase

3

115

9

0.45

68355/7.23

 P17475

α-1 antiproteinase

6

255

14

0.50

46281/5.70

 P85515

α-centractin

2

57

8

0.63

42703/6.19

Up-regulated proteins

 P04797

Glyceraldehyde-3-phosphate dehydrogenase

1

34

4

2.49

36095/8.14

 Q9WTT6

Guanine deaminase

6

312

16

1.54

51564/5.56

 P11598

Protein disulfide-isomerase A3

5

117

119

3.20

57052/5.88

 Q9Z0V6

Thioredoxin-dependent peroxide reductase. mitochondrial

2

78

9

2.09

28567/7.14

Accession number, protein name, number of matched peptides, proein score, percentage coverage, fold change (restricted/control) and theoretical molecular mass (Da) and pI of identified proteins

Table 6

Biological process classification of identified proteins of female rats

Protein name

Metabolic process

Down-regulated in restricted females

 Serine protease inhibitor A3K

Proteolysis

 Serine protease inhibitor A3L

 26S proteasome non-ATPase regulatory subunit 11

 Alpha-1-antiproteinase

 Transketolase

Carbohydrate, Amino acid, Lipidic

 2-oxoisovalerate dehydrogenase subunit beta, mitochondrial

 Perilipin-1

Lipidic

 Glutathione S-transferase theta-2

Protein

 L-lactate dehydrogenase B chain

Glycolysis, TCA

 Heterogeneous nuclear ribonucleoproteins A2/B1

Nucleotide

 Protein kinase C delta-binding protein

Transcription

 Staphylococcal nuclease domain-containing protein 1

 Ras-related protein Rap-1A

 

Up-regulated in restricted females

 Guanine deaminase

Purine

 Protein disulfide-isomerase A3

Protein folding

 Glyceraldehyde-3-phosphate dehydrogenase

Glycolysis

 Thioredoxin-dependent peroxide reductase, mitochondrial

 

Classification of proteins in metabolic process

In both males and females, some proteins were identified in 2 adjacent spots, what may possibly be attributed to the existence of either different isoforms or post-translational modifications of the protein.

Western blot analysis

A sub-set of selected proteins was analyzed by Western blotting to confirm the proteome results. Corroborating the male proteome result of a 40 % decrease in expression of 78 kDa glucose-regulated protein in the restricted males, the western blot analysis showed a 33 % decrease. The mitochondrial stress-70 protein showed a 48 % decrease in the proteome experiment and a 36 % decrease in the western blot experiment (Fig. 3).
Fig. 3

Western Blot analysis of 78 kDa glucose-regulated protein and Stress-70 protein. a 78 kDa glucose-regulated protein. N = 8 control; N = 9 restricted. b Stress 70 protein, mitochondrial. N = 10 controls; N = 12 restricted. *p < 0.05 vs. control

In the females, the proteome results showed a 68 % decrease of Glutathione S-transferase theta-2 in the restricted group while the western blot analysis showed a 25 % decrease (Fig. 4).
Fig. 4

Western Blot analysis of glutathione S-transferase theta 2. N = 8 controls; N = 9 restricted. * = p < 0.05 vs. control

Discussion

Male and female rats submitted to intrauterine growth restriction had normal food intake and body weight as adults, indicating catch-up growth, an adaptive mechanism against obesity in adult life [31, 32].

The restricted females, unlike the restricted males, showed increased fat pads weight, without overt peripheral insulin resistance, in accordance with a previous work from our laboratory [11]. This observation agrees with other reports showing the gender-dependency of the late consequences of rat maternal nutritional restriction [4, 5, 79]. In humans, early intrauterine undernutrition increased body mass index in 50 year-old women but not men [33].

The proteomic analysis of the adipose tissue of the males showed that IUGR caused alterations in the protein levels of 28 identified proteins. Levels of fatty acid synthase, enzyme of the de novo lipogenesis pathway [16], were increased by IUGR, in agreement with other reports [34, 35]. A study comparing lean and obese subjects found that increased fatty acid synthase gene expression was linked to visceral fat accumulation [36].

Prohibitin levels were down-regulated in the restricted males. This protein has been shown to attenuate insulin-stimulated oxidation of glucose and fatty acids in adipose tissue [37]. Over-expression of prohibitin in mice adipose tissue increased fat pads [38]. In contrast, knockdown of prohibitin in 3 T3-L1 pre-adipocytes increased oxidative stress due to impairment of mitochondrial function [39].

These protein expression alterations found in the restricted males, one favoring and the other counteracting lipid accumulation, may represent a pre-obese condition. Although the restricted males did not have augmented fat pads mass, they did show a tendency to hyperinsulinemia, suggesting that an increase in lipid synthesis could lead to obesity later in life.

The increased levels of proteasome subunit α type-3 suggest that IUGR caused stimulation of proteolysis in males. The adipose tissue has been shown to be an important site of proteolysis and to contribute to the circulating amino acids pool [40, 41]. Obese women showed a decreased rate of amino acids release from the tissue, in response to fasting [42].

In the adipose tissue of obese humans, levels of mitochondrial branched-chain-amino-acid aminotransferase were reportedly decreased from lean levels in the metabolically unhealthy but not in the healthy subset of obese subjects [43]. Here, tissue levels of mitochondrial branched-chain-amino-acid aminotransferase were increased in the restricted males, indicating that their metabolism was not affected at the same extent as that seen in unhealthy obesity.

IUGR up-regulated the levels of elongation factor 1-alpha 1, a GTPase that delivers aminoacyl–tRNAs to ribosomes during protein translation [44, 45]. This protein has been shown to interact with nascent proteins ubiquitinated during translation, facilitating their delivery to proteasome [46] and to be associated with stimulation of cell proliferation in cancer cells [47]. In kidneys of streptozotocin diabetic rats, increased expression of elongation factor-1A has been related to hypertrophy of the adipose organ and to diabetes-associated oxidative stress [48].

Obesity has recently been associated with increased levels of several amino acids in the visceral adipose tissue of humans [49]. Moreover, metabolomic analysis showed increased levels of phenylalanine, tryptophan and glutamate in the umbilical vein blood of IUGR neonates [50]. The increased levels of proteins related to proteolysis stimulation, as observed in the present study, may increase adipose tissue levels of amino acids. These may be converted to intermediates of the tricarboxylic acid (TCA) cycle. It is important to point out that, once entering the TCA cycle, these amino acids could be directed to either complete oxidation or generation of citrate [51], an important precursor for de novo lipogenesis. It is thus reasonable to suggest that proteolysis stimulation in the restricted males may provide amino acids for metabolic reactions in the tissue, rather than for release. A recent review has indicated that impairment of TCA cycle metabolites by IUGR could be an important biomarker of this condition [52].

Cytosolic malate dehydrogenase levels were up-regulated in the restricted males. This enzyme is active in the malate/aspartate shuttle, where it catalyzes the reduction of oxaloacetate to malate, using NADH. Malate enters mitochondria and is oxidized to oxaloacetate by mitochondrial malate dehydrogenase, with production of NADH. This shuttle not only channels the NADH produced during glycolysis to ATP production but also maintains the cytosolic NAD+/NADH ratio, essential for the oxidative metabolism of glucose [5355]. Increased levels of mitochondrial malate dehydrogenase have been reported in pancreatic islets of adult rats with IUGR. However, ATP levels were not altered, which was attributed to the concomitant decrease of ATP synthase subunit 6 levels [55]. In the present study, ATP synthase subunit alpha was up-regulated, indicating that ATP production could be increased.

Some proteins down-regulated by IUGR in males are related to inflammation and cellular stress. Murinoglobulin-1 is a serino-protease inhibitor [56] that plays a protective role in the inflammatory response. Hemopexin is a positive acute-phase reactant that plays a protective role in lipid peroxidation through its heme binding effect [57], its levels being negatively associated with the severity of chronic sepsis [58]. In diet-induced obese mice, up-regulation of serum hemopexin levels has been suggested to represent a dysfunctional response in this chronic inflammatory condition [59].

The 78 kDa glucose-regulated protein is related to proper protein folding, protecting the cell from endoplasmic reticulum stress [60, 61], which has been described to link obesity to insulin resistance [61]. Obese mice overexpressing 78 kDa glucose-regulated protein in pancreas were protected against endoplasmic reticulum stress and had improvement of insulin sensitivity [62]. Heat shock protein HSP 90-beta is an important chaperone whose levels reportedly increase in obese humans, playing a role in mitigating the inflammatory stress present in obesity [63, 64]. Taken together, these protein alterations indicate that the restricted males presented impairment of anti-inflammatory reactions in the adipose tissue.

Overall, the results found in the male rats indicate that, even though the restricted males did not have augmented fat pads or glucose intolerance, the alterations in adipose tissue metabolism point to a tendency to develop obesity.

In the females, IUGR affected the glycolysis/gluconeogenesis pathway. L-lactate dehydrogenase B was down-regulated in the restricted females, indicating low production of lactate from pyruvate. Due to its low blood supply, the adipose tissue produces considerable amounts of lactate, which can serve either as precursor to energy production or fatty acid synthesis [65, 66] or be released to the systemic circulation [67], even in normoxia conditions [68]. Adipose tissue lactate production has been shown to correlate with lactate dehydrogenase activity, both under normal and cafeteria diet feeding, and suggested to contribute to glycemic control, through consumption of excess circulating glucose [69].

Glyceraldehyde-3-phosphate dehydrogenase, the enzyme catalyzing the reversible conversion of glyceraldehyde-3-phosphate to 1,3 bisphosphoglycerate and NADH, was up-regulated in the restricted females. Stimulation of glyceraldehyde-3-phosphate dehydrogenase has been reported in pre-obese, normoinsulinemic, Zucker rats [70]. Maternal peri-conceptional overnutrition, but not food restriction, increased fat mass of postnatal female lambs and glyceraldehyde-3-phosphate dehydrogenase gene expression correlated positively with perirenal fat amount [71].

Transketolase was down-regulated in the restricted females. This enzyme catalyzes the formation of glyceraldehyde-3-phospate in the non-oxidative branch of the pentose phosphate pathway, in which ribose is re-converted to glucose. Moreover, the pentose phosphate pathway generates NADPH for lipid synthesis. In obese individuals, decreased activity of the lipogenic pathway, with down-regulation of transketolase, has been interpreted as a mechanism aimed at reducing the growth of adipose tissue [72].

Reduced levels of mitochondrial branched-chain-amino-acid aminotransferase and mitochondrial 2-oxoisovalerate dehydrogenase (also known as branched-chain alpha-keto acid dehydrogenase E1), enzymes participating in the pathway of degradation of branched-chain amino acids, were found in the subcutaneous adipose tissue of unhealthy obese humans but not in the healthy obese subset [43]. Here, the latter enzyme was down-regulated in the restricted females. It is possible to suggest that the metabolism of branched-chain-amino-acids in the adipose tissue of the restricted females resembled that found in obesity associated with metabolic derangements. This contrasts with the result in the restricted males, in which mitochondrial branched-chain-amino-acid aminotransferase was increased.

The restricted females also showed down-regulation of perilipin-1, an enzyme active in lipid droplet formation [73] and inversely correlated with adipocyte size and basal lipolysis [74]. Perilipin gene suppression increased basal lipolysis and prevented high-fat diet obesity in mice [75].

Glutathione S-transferase theta-2 was down-regulated in the restricted females. This protein is part of the antioxidant enzymes family, which catalyzes the conjugation of glutathione to a wide variety of compounds. Decreased glutathione or glutathione-S transferase levels have been linked to diabetes, due to its role in antioxidant pathways [76, 77]. On the other hand, high levels of glutathione-S transferase P in obese subjects activated inflammatory pathways and endoplasmic reticulum stress [78].

Other proteins related to antioxidant pathways were up-regulated by IUGR in the females. Protein disulfide-isomerase A3 is a thiol-disulfide oxidoreductase present in the endoplasmic reticulum and it catalyzes the formation, breakdown and rearrangement of disulfide bonds [79]. Increased levels of protein disulfide-isomerase A3 in the adipose tissue of obese subjects have been suggested to activate inflammatory pathway and endoplasmic reticulum stress [79]. Mitochondrial thioredoxin-dependent peroxide reductase regulates H2O2 levels, protecting the cell from the toxicity resulting from its accumulation [80], and depletion of this protein accelerated apoptosis [81].

A protein down-regulated in the restricted females, α-1-antiproteinase, also known as serpin A1 and α1-antitrypsin, is a serine protease inhibitor with anti-inflammatory effects. It caused inhibition of lipopolysaccharide-mediated activation of in vitro human monocytes [82] and inhibited lung neutrophil chemotaxis [8385]. Inhalation of α-1-antiproteinase decreased protein levels of IL-1β and IL-8 [86] while addition of purified plasma α-1-antiproteinase to pancreatic β-cells in vitro inhibited cytokine-induced apoptosis [87]. Alpha-1-antiproteinase gene therapy prevented the development of type 1 diabetes in non-obese mice [88]. Decreased levels of α-1-antiproteinase was reported by proteomic analysis of adipose tissue of women with gestational diabetes mellitus [89] impair the protection against inflammation and oxidative stress, compensatory mechanisms were recruited in the restricted females.

Conclusions

In the restricted males, the high levels of proteasome subunit α type 3, branched-chain-amino-acid aminotransferase and elongation 1- alpha 1 indicate increased proteolysis rate in the adipose tissue. High tissue levels of amino acids could generate lipogenesis precursors, a suggestion supported by the high levels of fatty acid synthase. The increased levels of cytosolic malate dehydrogenase and ATP synthase subunit alpha may favor ATP production. These results indicate that, in the restricted males, the alterations in protein expression induced by IUGR pointed to a metabolic status favoring the development of obesity.

In the restricted females, the decreased levels of perilipin-1 are indicative of increased lipolysis while the low levels of mitochondrial branched-chain alpha-keto acid dehydrogenase E1 indicate low proteolysis rate. The low levels of transketolase could represent low activity of the pentose phosphate pathway and, consequently, decreased lipogenesis rate. Down-regulation of L-lactate dehydrogenase may lead to impairment of glycemic control. These alterations point to a metabolic status of established obesity in the restricted females. In both genders, the protein variations indicated impairment of pathways involved in the responses to oxidative stress and inflammation (Fig. 5).
Fig. 5

Diagram of the suggested pathways modified by IUGR in the retroperitoneal adipose tissue of male (a) and female (b) rats

Methods

Rats

Wistar rats were cared for in accordance with the guidelines of the committee on animal research ethics of the Federal University of São Paulo (approval 486691). Three months-old rats were mated and the first day of pregnancy was determined by examination of vaginal smears for the presence of sperm. From day 1 of pregnancy, the dams were randomly assigned to be a control or a restricted dam. The control dams were fed ad libitum throughout pregnancy and lactation. The restricted dams received only 50 % of control intake during the whole pregnancy and were fed ad libitum during lactation. On the day of delivery, the pups were adjusted to eight per dam.

After weaning, the male and female offspring from control and restricted dams were housed four/five per cage and fed ad libitum until 4 months of age. The food provided to dams and offspring consisted of standard rat chow (Nuvital Nutrients, Columbo, PR, Brazil) containing (w/w) 4.5 % fat, 23 % protein, and 33 % carbohydrate, with 2.7 kcal/g, as determined at the Bromatology Division of the Federal University of São Paulo. All animals were maintained in controlled conditions of lighting (12-h light/12-h dark cycle, lights off at 18:00 h) and temperature (24 ± 1 °C) and had free access to water throughout the experimental period.

The numbers of animals used in the study were 16 male and 14 female controls and 17 male and 14 female restricted rats.

Weight gain, weight of white adipose tissue and blood and tissue measurements

Food intake and body weight were measured once a week since weaning. At 4 months of age, the animals were killed by decapitation. The retroperitoneal adipose tissue was rapidly removed, weighed and frozen in liquid nitrogen. The tissue was stored at - 80 °C until analysis. The gonadal and mesenteric white adipose tissues were dissected and weighed.

Trunk blood was centrifuged and the serum stored at - 80 °C. Glucose analysis was performed by the glucose oxidase method, using a commercially available kit with detection limit of 0.32 mg/dL (Glucose Pap Liquiform, Labtest Diagnostica, São Paulo, Brazil). Triglycerides levels were determined using a commercially available kit with detection limit of 0.82 mg/dL (Labtest Diagnostica, São Paulo, Brazil). Insulin, corticosterone, adiponectin, TNF-α and IL-1β levels were measured by multiplex kit (Millipore, Bedford, MA, USA). The measurements of TNF-α, IL-10 and IL-6 in tissue were performed by Elisa (Millipore, Bedford, MA, USA).

Proteome analysis

Sample preparation

An aliquot of 700 mg of retroperitoneal adipose tissue was homogenized in 1 ml of extraction buffer (7 M urea, 2 M thiourea, 4 % (w/v) CHAPS, 0.5 % (v/v) Triton X-100) containing complete Mini Protease Inhibitor Cocktail Tablets (Roche Diagnostics, Germany), added immediately before use. Sample lysates were centrifuged (19,000 g/30 min.) and supernatants stored at - 80 °C until analysis.

Protein assay

Protein concentration of supernatants was determined using 2-D Quant Kit (GE Healthcare, Pittsburgh, USA) and bovine albumin as standard, according to manufacturer’s recommendations.

Protein precipitation

Aliquots of 900 μg of protein were precipitated with a solution of 35 % KCl, 44 % chloroform, and 21 % methanol (v/v). The mixture was homogenized and centrifuged at 19,000 g and 4 °C for 15 min. The pellet was air-dried at room temperature.

Two-dimensional gel electrophoresis and image analysis

For isoelectric focusing (IEF), the pellet was dissolved in 500 μL of rehydration buffer (7 M urea, 2 M thiourea, 4 % (w/v) CHAPS, 0.5 % (v/v) Triton X-100, 100 mM DTT, 0.2 % (v/v) IPG Buffer pH 3–10, and traces of Bromofenol blue). IEF was carried out on a Protean IEF cell (Bio-Rad, CA, USA) using immobiline dry strips (18 cm linear gradient, pH 3–10) previously rehydrated for 12–14 h. IEF was performed with the current limit set at 50 mA per IPG strip with the following conditions at 18 °C: 100 V for 30 min, 250 V for 2 h, 500 V for 30 min, 1000 V for 30 min, 2000 V for 30 min, 4000 V for 1 h, 8000 V for 1 h followed by 8000 V until 30000 Vh.

After focusing, strips were equilibrated for 25 min in buffer containing 6 M urea, 50 mM Trisma base pH 8.8, 34 % (v/v) glycerol, 2 % (w/v) SDS, and 1 % (w/v) DDT, followed by an additional 25 min in the same buffer containing 2.5 % (w/v) iodoacetamide instead of DTT. Strips were then loaded onto 12 % SDS- polyacrylamide gels. After running in Protean II Multi-Cell (Bio-Rad, CA, USA), at 50 mA per gel for 6 h, the gels were stained for 48 h with Coomassie Blue G-250 (Bio-Rad, CA, USA). Stained gels were scanned (GS-710 Calibrated Imaging Densitometer) and analyzed using PDQuest Image Analysis Software version 7.2 (Bio-Rad, CA, USA).

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry

The selected spots were manually excised, distained and digested. The spots were excised and distained in 50 % methanol and 5 % acetic acid overnight. The excised spots were treated with 25 mM ammonium bicarbonate and 50 % acetonitrile (1:1) and dried in SpeedVac. To the dried spots, 10 mM DTT was added and incubated for 1 h at 56 °C, followed by 55 mM IAA for 45 min on the dark. The spots were dehydrated with 25 mM ammonium bicarbonate followed by 25 mM ammonium bicarbonate with 50 % acetonitrile and dried in SpeedVac. Digestion was performed overnight with 15 ng of trypsin (Promega, WI, USA) in 25 mM ammonium bicarbonate, at 37 ° C. Digested samples were desalted using C18 Zip Tips (Millipore, Bedford, MA, USA). Two microliters of sample were applied on the spectrometer plate and air-dried at room temperature. The matrix solution (10 mg/mL α-cyano-4 hydroxycinnamic acid in 70 % acetonitrile/0.1 % trifluoroacetic acid) was applied on the spectrometer plate and air-dried at room temperature.

MALDI-TOF/TOF MS was performed using an Axima Performance ToF-ToF, (Kratos-Shimadzu Biotech, Manchester, UK) mass spectrometer. The instrument was externally calibrated with [M + H]+ ions of bradykinin (1–7 fragment, 757.4 Da), human angiotensin II (1046.54 Da), P14R synthetic peptide (1533.86 Da), and human ACTH (18–39 fragment, 2465.20 Da). Following MALDI MS analysis, MALDI MS/MS was performed on the 7 most abundant ions from each spot.

MASCOT (Matrix Science, UK) server was used to search Swiss-Prot protein database (http://www.matrixscience.com). The following parameters were used in this search: no restrictions on protein molecular weight, trypsin digest with one missing cleavage, monoisotopic mass, taxonomy limited to Rattus, carbamidomethylation of cysteine as fixed modification, possible oxidation of methionine and tryptophan, peptide mass tolerance of 0.5 Da, fragment mass tolerance of 0.8 Da, and peptide charge +1. False discovery rate (FDR) assessment was estimated using Mascot decoy database approach and only proteins identified with 0 % FDR were included in the results. Protein matching probabilities were determined using MASCOT protein scores, with identification confidence indicated by the number of matching and the coverage of protein sequence by the matching peptides. The presence of at least one peptide with significant ion score was required for positive protein identification. Only statistically significant MASCOT score results (p < 0.05) were included in the analysis.

The identified proteins were classified in Panther (http://www.pantherdb.org/) according to biological process.

Western Blot analysis

A sub-set of adipose tissue samples was used in western blot experiments. A 700 mg aliquot was homogenized in 1.0 ml of solubilization buffer (10 mM EDTA, 100 mM Tris pH 7.5, 10 mM sodium pyrophosphate, 100 mM sodium fluoride, 10 mM sodium orthovanadate, 2 mM PMSF, aprotinin 2 μg/mL, and 1 % Triton X-100). Insoluble material was removed by centrifugation (19,000 g at 4 ° C for 40 min.). The supernatant was collected and one aliquot was separated for protein concentration determination. Tissue extracts were denatured by boiling for 5 min in Laemmli buffer [90] containing 100 mM DTT. The protein concentration was determined by colorimetric method (BCA Protein Assay, Bioagency Biotecnologia, Brazil).

Subsequently, protein extracts (100 μg) were resolved in 12 % SDS polyacrylamide gels and transferred to nitrocellulose membranes using a semi-dry transfer system (Bio-Rad, CA, USA). Non-specific binding sites were blocked for 2 h in 1 % bovine serum albumin. The nitrocellulose membranes were then incubated overnight with primary antibody and for 1 h with the appropriate secondary antibody conjugated with horseradish peroxidase. The quantitative analysis was performed by densitometry using Scion Image software (Scion Corporation, Frederick, MD, USA).

The results were expressed in arbitrary units, as percentage changes in relation to the control group. For evaluation of protein loading, all membranes were stripped and reblotted with anti-β-tubulin (for male) and anti-β-actin (for female) primary antibody. The antibodies against 78 kDa glucose regulated protein (1:1000; ab53068), mitochondrial stress 70 protein (1:1000; ab106654), and glutathione S-transferase theta-2 (1:2500; ab102045) were obtained from ABCAM (Cambridge, UK). The antibody against β-tubulin (1:5000; #2146S) was purchased from Cell Signaling (Danvers, MA, USA). The antibody against β-actin (1:1000; sc-130657) was purchased from Santa Cruz (Dallas, TX, USA).

Statistical analysis

The data are expressed as mean ± SEM. Comparisons between groups (control and restricted) were performed by Student t test. Statistical significance was set at p < 0.05.

Abbreviation

2DE: 

two-dimensional gel electrophoresis

ATP: 

adenosine triphosphate

BW: 

body weight

FC: 

Fold change

IL: 

interleukin

IUGR: 

intrauterine growth restriction

MW: 

Molecular weight

NADH: 

Nicotinamide adenine dinucleotide

TCA: 

trycarboxilic acid cycle

TNF-α: 

tumor nuclear factor alpha

Declarations

Acknowledgements

This research was supported by grants from the Brazilian agencies: State of São Paulo Research Foundation (FAPESP), National Council for Scientific and Technological Development (CNPq), and Coordination for the Enhancement of Higher Education Personnel (CAPES).

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Authors’ Affiliations

(1)
Departamento de Fisiologia, Universidade Federal de São Paulo
(2)
Centro de Química de Proteínas – Hemocentro, Universidade de São Paulo

References

  1. Langley-Evans SC. Nutritional programming of disease: unravelling the mechanism. J Anat. 2009;215:36–51.PubMed CentralView ArticlePubMedGoogle Scholar
  2. Breton C. The hypothalamus-adipose axis is a key target of developmental programming by maternal nutrition manipulation. J Endocrinol. 2013;216(2):R19–31.View ArticlePubMedGoogle Scholar
  3. Hajj NE, Schneider E, Lehnen H, Haaf T. Epigenetics and life-long consequences of an adverse nutritional and diabetic intrauterine environment. Reproduction. 2014;148:R111–20.PubMed CentralView ArticlePubMedGoogle Scholar
  4. Aiken CE, Ozanne SE. Sex differences in developmental programming models. Reproduction. 2013;145:R1–13.View ArticlePubMedGoogle Scholar
  5. Picó C, Palou M, Priego T, Sánchez J, Palou A. Metabolic programming of obesity by energy restriction during the perinatal period: different outcomes depending on gender and period, type and severity of restriction. Front Physiol. 2012;3:1–14.View ArticleGoogle Scholar
  6. Fisher RE, Steele M, Karrow NA. Fetal programming of the neuroendocrine-immune system and metabolic disease. J Preg. 2012. doi:10.1155/2012/792934.Google Scholar
  7. Howie GJ, Sloboda DM, Vickers MH. Maternal undernutrition during critical windows of development results in differential and sex-specific effects on postnatal adiposity and related metabolic profiles in adult rat offspring. Br J Nutr. 2012;108:298–307.View ArticlePubMedGoogle Scholar
  8. Manuel-Apolinar L, Rocha L, Damasio L, Tesoro-Cruz E, Zarate A. Role of prenatal undernutrition in the expression of serotonin, dopamine and leptin receptor in adult mice: implication of food intake. Mol Med Reports. 2014;9:407–12.Google Scholar
  9. Anguita RM, Sigulem DM, Sawaya AL. Intrauterine food restriction is associated with obesity in young rats. J Nutr. 1993;123:1421–8.PubMedGoogle Scholar
  10. Porto LCJ, Sardinha FLC, Telles MM, Guimarães RB, Albuquerque KT, Andrade IS, et al. Impairment of the serotonergic control of feeding in adults female rats expose to intra-uterine malnutrition. Br J Nutr. 2009;101:1255–61.View ArticlePubMedGoogle Scholar
  11. Sardinha FLC, Telles MM, Albuquerque KT, Oyama LM, Guimarães PAMP, Santos OFP, et al. Gender difference in the effect of intrauterine malnutrition on the central anorexigenic action of insulin in adult rats. Nutrition. 2006;22:1152–61.View ArticlePubMedGoogle Scholar
  12. Vickers MH, Breier BH, Cutfield WS, Hofman PL, Gluckman PD. Fetal origins of hyperphagia, obesity, and hypertension and postnatal amplification by hypercaloric nutrition. Am J Physiol. 2000;279:E83–7.Google Scholar
  13. Bieswal F, Ahn M, Reusens B, Holvoet P, Raes M, Rees WD, et al. The importance of catch-up growth after early malnutrition for the programming of obesity in male rat. Obesity. 2006;14:1330–43.View ArticlePubMedGoogle Scholar
  14. Delahaye F, Lukaszewski M-A, Wattez J-S, Cisse O, Dutriez-Casteloot I, Fajardy I, et al. Maternal perinatal undernutrition programs a “brown-like” phenotype of gonadal white fat in male rat at weaning. Am J Physiol Regul Integr Comp Physiol. 2010;299:R101–10.View ArticlePubMedGoogle Scholar
  15. Alexandre-Goubau M-CF, Courant F, Le-Gall G, Moyon T, Darmaun D, Parnet P, et al. Offspring metabolomic response to maternal protein restriction in a rat model of intrauterine growth restriction (IUGR). J Proteome Res. 2011;10:3292–302.View ArticleGoogle Scholar
  16. Proença ARG, Sertié RAL, Oliveira AC, Campaña AB, Caminhotto RO, Chimin P, et al. New concepts in White adipose tissue physiology. Braz J Med Biol Res. 2014;47(3):192–205.PubMed CentralView ArticlePubMedGoogle Scholar
  17. Belfiore F, Rabuazzo AM, Napoli E, Borzi V, Vecchio LL. Enzymes of glucose metabolism and of citrate cleavage pathway in adipose tissue of normal and diabetes subjects. Diabetes. 1975;24:865–73.View ArticlePubMedGoogle Scholar
  18. Langin D. Adipose tissue lipolysis as a metabolic pathway to define pharmacological strategies against obesity and metabolic syndrome. Pharmacol Res. 2006;53:482–91.View ArticlePubMedGoogle Scholar
  19. Dessì A, Pravettoni C, Marincola FC, Schirru A, Fanos V. The biomarkers of fetal growth in intrauterine growth retardation and large for gestational age cases: from adipocytokines to a metabolomic all-in-one tool. Expert Rev Proteomics. 2015;12(3):309–16.View ArticlePubMedGoogle Scholar
  20. Kershaw EE, Flier JS. Adipose tissue as an endocrine organ. J Clin Endocrinol Metab. 2004;89(6):2548–56.View ArticlePubMedGoogle Scholar
  21. Nascimento CMO, Ribeiro EB, Oyama LM. Metabolism and secretory function of white adipose tissue: effect of dietary fat. Anais Acad Bras Cienc. 2009;81(3):453–66.View ArticleGoogle Scholar
  22. Briana DD, Malamitsi-Puchner A. Intrauterine growth restriction and adult disease: the role of adipocytokines. Eur J Endocrinol. 2009;160:337–47.View ArticlePubMedGoogle Scholar
  23. Ibáñez L, Sebastiani G, Lopez-Bermejo A, Díaz M, Gómez-Roig MD, de Zegher F. Gender specificity of body adiposity and circulating adiponectin, visfatin, insulin, and insulin growth factor-I at term birth: relation to prenatal growth. J Clin Endocrinol Metab. 2008;93(7):2774–8.View ArticlePubMedGoogle Scholar
  24. Fuchs D, Winkelmann I, Johnson IT, Mariman E, Wenzel U, Daniel H. Proteomics in nutrition research: principles, technologies and applications. Br J Nutr. 2005;94:302–14.View ArticlePubMedGoogle Scholar
  25. Wang J, Li D, Dangott LJ, Wu G. Proteomics and its role in nutrition research. J Nutr. 2006;136:1759–62.PubMedGoogle Scholar
  26. Roepstorff P. Mass spectrometry-based proteomics, background, status and future needs. Protein Cell. 2012;3(9):641–7.View ArticlePubMedGoogle Scholar
  27. Silva TS, Richard N, Dias JP, Rodrigues PM. Data visualization and futures selection methods in gel-based proteomics. Cur Protein Pep Sci. 2014;15:4–22.View ArticleGoogle Scholar
  28. You Y-A, Lee JH, Kwon EJ, Yoo JY, Kwon W-S, Pang M-G, Kim YJ. Proteomic analysis of one-carbon metabolism-related marker in liver of rat offspring. Mol Cel Proteomics. 2015. Paper in press.Google Scholar
  29. Sarr O, Louveau I, Kalbe C, Metges CC, Rehfeldt C, Gondret F. Prenatal exposure to maternal low or high protein diets induces modest changes in the adipose tissue proteome of newborn piglets. J Anim Sci. 2010;88:1626–41.View ArticlePubMedGoogle Scholar
  30. Ruis-Gonzáles MD, Cañete MD, Gómez-Chaparro JL, Abril N, Cañete R, López-Barea J. Alteration of protein expression in serum of infants with intrauterine growth restriction and different gestational age. J Proteomics. 2015;119:169–82.View ArticleGoogle Scholar
  31. Alexandre-Goubau M-CF, Bailly E, Moyon TL, Grit IC, Coupé B, Drean GL, et al. Postnatal growth velocity modulates alterations of proteins involved in metabolism and neuronal plasticity in neonatal hypothalamus in rats born with intrauterine growth restriction. J Nutr Biochem. 2012;23:140–52.View ArticleGoogle Scholar
  32. Fabricius-Bjerre S, Jensen RB, Faerch K, Larsen T, Molgaard C, Michaelsen KF, et al. Impact of birth weight and early infant weight gain on insulin resistance and associated cardiovascular risk factors in adolescence. Plos One. 2011;6(6):e20595. doi:10.1371/journal.pone.0020595.PubMed CentralView ArticlePubMedGoogle Scholar
  33. Ravelli ACJ, van der Meuelen JHP, Osmond C, Barker DJP, Bleker OP. Obesity at the age 50 y in men and women exposed to famine prenatally. Am J Clin Nutr. 1999;70:811–6.PubMedGoogle Scholar
  34. Desai M, Han G, Ferelli M, Kallichanda N, Lane RH. Programmed upregulation of adipogenic transcriptions factors in intrauterine growth-restricted offspring. Reprod Sci. 2008;15(8):785–96.PubMed CentralView ArticlePubMedGoogle Scholar
  35. Lukaszewski M-A, Mayer S, Fajardy I, Delahaye F, Dutriez-Casteloot I, Montel V, et al. Maternal prenatal undernutrition programs adipose tissue gene expression in adult male rat offspring under high-fat diet. Am J Physiol Endocrinol Metab. 2011;301:E548–59.View ArticlePubMedGoogle Scholar
  36. Berndt J, Kovacs P, Ruschke K, Klöting N, Fasshauer M, Schön MR, et al. Fatty acid synthase gene expression in humana adipose tissue: assossiation with obesity and type 2 diabetes. Diabetologia. 2007;50:1472–80.View ArticlePubMedGoogle Scholar
  37. Vessal M, Mishra S, Moulik S, Murphy LJ. Prohibitin attenuates insulin-stimulated glucose and fatty acid oxidation in adipose tissue by inhibition of pyruvate carboxylase. FEBS J. 2006;273:568–76.View ArticlePubMedGoogle Scholar
  38. Ande SR, Nguyen KH, Padilla-Meier GP, Wahida W, Nyomba BLG, Mishra S. Prohibitin overexpression in adipocytes induces mitochondrial biogenesis, leads to obesity development, and affects glucose homeostasis in a sex-specific manner. Diabetes. 2014;63:3734–41.View ArticlePubMedGoogle Scholar
  39. Liu D, Lin Y, Kang T, Huang B, Xu W, Garcia-Barro M, et al. Miochondrial dysfunction and adipogenic reduction by prohibitin silencing in 3 T3-L1 cells. PLoS One. 2012;7(3):e34315. doi:10.1371/journal.pone.0034315.PubMed CentralView ArticlePubMedGoogle Scholar
  40. Herman MA, She P, Peroni OD, Lynch CJ, Kahn BB. Adipose tissue branched chain amino acid (BCAA) metabolism modulates circulating BCAA levels. J Biol Chem. 2010;285(15):11348–56.PubMed CentralView ArticlePubMedGoogle Scholar
  41. Kowalski TJ, Wu G, Watford M. Rat adipose tissue amino acid metabolism in vivo as assessed by microdialysis and arteriovenous techniques. Am J Physiol. 1997;273(3):E613–22.PubMedGoogle Scholar
  42. Patterson BW, Horowitz JF, Wu G, Watford M, Coppack SW, Klein S. Regional muscle and adipose tissue amino acid metabolism in lean and obese women. Am J Physiol Endocrinol Metab. 2011;282:E931–6.View ArticleGoogle Scholar
  43. Badoud F, Lam KP, DiBattista A, Perreault M, Zulyniak MA, Cattrysse B, et al. Serum and adipose tissue amino acid homeostasis in the metabolically healthy obese. J Proteome Res. 2014;13:3455–66.View ArticlePubMedGoogle Scholar
  44. Hershey JW. Translational control in mammalian cell. Annu Rev Biochem. 1991;60:717–55.View ArticlePubMedGoogle Scholar
  45. Thornton S, Anand N, Purcell D, Lee J. Not just for housekeeping: protein initiation and elongation factors in cell growth and tumorigenesis. J Mol Med. 2003;81:536–48.View ArticlePubMedGoogle Scholar
  46. Chuang S-M, Chen L, Lambertson D, Anand M, Kinzy TG, Madura K. Proteasome-mediated degradation of cotranslationally damage proteins involves translation elongation factor 1A. Mol Cell Biol. 2005;25(1):403–13.PubMed CentralView ArticlePubMedGoogle Scholar
  47. Al-Maghrebi M, Anin JT, Olalu AA. Up-regulation of eukaryotic elongation factor 1 subunits in breast carcinoma. Anticancer Res. 2005;25:2573–8.PubMedGoogle Scholar
  48. Al-Maghrebi M, Cojocel C, Thompson MS. Regulation of elengation factor 1 expression by vitamin E in diabetic rat kidney. Mol Cell Biochem. 2005;273:177–83.View ArticlePubMedGoogle Scholar
  49. Hanzu FA, Vinaixa M, Papageourgiou A, Párrizas M, Correig X, Delgado S, et al. Obesity rather than regional fat depots marks the metabolomic pattern of adipose tissue: an untargeted metabolomic approach. Obesity. 2014;22:698–704.View ArticlePubMedGoogle Scholar
  50. Favretto D, Cosmi E, Ragazzi E, Visentin S, Tucci M, Fais P, et al. Cord blood metabolomic profiling in intrauterine growth restriction. Anal Bioanal Chem. 2012;402:1109–21.View ArticlePubMedGoogle Scholar
  51. Lee S-M, Dho SH, Ju S-K, Maeng J-S, Kim J-Y, Kwon K-S. Cytosolic malate dehydrogenate regulates senescence in human fybroblasts. Biogereontology. 2012;13:525–36.View ArticleGoogle Scholar
  52. Dessì A, Puddu M, Ottonello G, Fanos V. Metabolomics and fetal-neonatal nutrition: between “not enough” and “too much”. Molecules. 2013;18:11724–32.View ArticlePubMedGoogle Scholar
  53. Mali Y, Zisapels N. Gain of interaction of ALS-linked G93A superoxide dismutase with cytosolic malate dehydrogenase. Neurobiol Dis. 2008;32:133–41.View ArticlePubMedGoogle Scholar
  54. Minárik P, Tomásková N, Kollárová M, Antalík M. Malate dehydrogenase – structure and function. Gen Physiol Biophys. 2002;21:257–65.PubMedGoogle Scholar
  55. Theys N, Ahn M-T, Bouckenooghe T, Reusens B, Remacle C. Maternal malnutrition programs pancreatic islet mitochondrial dysfunction in the adult offspring. J Nutr Biochem. 2011;22:985–94.View ArticlePubMedGoogle Scholar
  56. Saito A, Shinohara H. Rat plasma murinoglobulin: isolation, characterization, and comparison with rat α-1- and α-2-macroglobulins. J Biochem. 1985;98:501–16.PubMedGoogle Scholar
  57. Tolosano E, Altruda F. Hemopexin: structure, function, and regulation. DNA Cell Biol. 2002;21(4):297–306.View ArticlePubMedGoogle Scholar
  58. Jung JY, Kwak YH, Kim KS, Kwon WY, Suh GJ. Change of hemopexin level is associated with the severity of sepsis in endotoxemic rat model and the outcome of septic patients. J Crit Care. 2015. doi:10.1016/j.jcrc.2014.12.009.PubMedGoogle Scholar
  59. Gianazza E, Sensi C, Eberini I, Gilardi F, Giudici M, Crestani M. Inflammatory serum proteome pattern in mice fed a high-fat diet. Amino Acids. 2013;44:1001–8.View ArticlePubMedGoogle Scholar
  60. Walter P, Ron D. The unfolded protein response: to stress pathway to homeostatic regulation. Science. 2011;334:1081–6.View ArticlePubMedGoogle Scholar
  61. Özcan U, Cao Q, Yilmaz E, Lee A-H, Iwakoshi NN, Özdelen E, et al. Endoplasmic reticulum stress links obesity, insulin action, and type 2 diabetes. Science. 2004;306:457–61.View ArticlePubMedGoogle Scholar
  62. Teodoro-Morrison T, Schuiki I, Zhang L, Belsham DD, Volchuk A. GRP78 overproduction in pancreatic beta cells protects against high-fat-diet-induced diabetes in mice. Diabetologia. 2013;56:1057–67.View ArticlePubMedGoogle Scholar
  63. Lanneau D, Brunet M, Frisan E, Solary E, Fontenay M, Garrido C. Heat shock proteins : essential proteins for apoptosis regulation. J Cell Mol Med. 2008;12(3):743–61.PubMed CentralView ArticlePubMedGoogle Scholar
  64. Tiss A, Khadir A, Abubaker J, Abu-Farha M, Al-Khairi I, Cherian P, et al. Immunohistochemical profiling of the heat shock response in obese non-diabetic subjects revealed impairment expression of heat shock proteins in the adipose tissue. Lipids Health Dis. 2014. doi:10.1186/1476-511X-13-106.PubMed CentralPubMedGoogle Scholar
  65. Saggerson ED, McAllister TWJ, Bath HS. Lipogenesis in rat brown adipocytes – effects of insulin and noradrenaline, contributions from glucose and lactate as precursors and comparisons with white adipose tissue. Biocem J. 1988;251:701–9.View ArticleGoogle Scholar
  66. O'Hea EK, Leveille G. Significance of adipose tissue and liver as sites of fatty acid synthesis in the pig and the efficiency of utilization of various substrates for lipogenesis. J Nutr. 1969;99:338–44.PubMedGoogle Scholar
  67. van Hall G. Lactate kinects in human tissues at rest and during exercise. Acta Physiol. 2010;199:499–508.View ArticleGoogle Scholar
  68. Sabbater D, Arriarán S, Romero MM, Agnelli S, Remesar X, Fernández-López JA, et al. Cultured 3 T3-L1 adipocytes dispose of excess medium glucose as lactate under abundant oxygen availability. Sci Rep. 2014;4:3663. doi:10.1038/srep03663.Google Scholar
  69. Arriaran S, Agnelli S, Sabater D, Remesar X, Fernádez-López JA, Alemany M. Evidences of basal lactate production in the main white adipose tissue sites of rats. Effect of sex and a cafeteria diet. PLoS One. 2015;10(3):e0119572. doi:10.1371/journal.pone.0119572.PubMed CentralView ArticlePubMedGoogle Scholar
  70. Dugail I, Quignard-Boulange A, Bazin R, Le Liepvre X. Adipo-tissue-specific increase in glyceraldehyde-3-phosphate dehydrogenase activity and mRNA amounts in suckling pre-obese Zucker rats. Biochem J. 1988;254:483–7.PubMed CentralView ArticlePubMedGoogle Scholar
  71. Rattanatray L, MacLaughlin SM, Kleemann DO, Walker SK, Muhlhausler BS, McMillen IC. Impact of maternal periconceptional overnutrion on fat mass and expression of adipogenic and lipogenic genes in visceral and subcutaneous fat depots in the postnatal lamb. Endocrinology. 2010;151(11):5195–205.View ArticlePubMedGoogle Scholar
  72. Pérez-Pérez R, García-Santos E, Ortega-Delgado FJ, López JA, Camafeita E, Ricart W, et al. Attenuated metabolism is a hallmark of obesity as revealed by comparative proteomic analysis of human omental adipose tissue. J Proteomics. 2012;75:783–95.View ArticlePubMedGoogle Scholar
  73. Brasaemle DL. The perilipin family of structural lipid droplet proteins: stabilization of lipid droplets and control of lipolysis. J Lipid Res. 2007;48:2547–59.View ArticlePubMedGoogle Scholar
  74. Ray H, Pinteur C, Frering V, Beylot M, Large V. Depot-specific differences in perilipin and hormone-sensitive lipase expression in lean and obese. Lipid Health Dis. 2009;8(58). doi:10.1186/1476-511X-8-58.
  75. Tansey J, Sztalryd C, Gruia-Gray J, Roush DL, Zee JV, Gavrilova O, et al. Perilipin ablation results in a lean mouse with aberrant adipocyte lipolysis, enhanced leptin production, and esistance to diet-induced obesity. Proc Natl Acad Sci. 2001;98:6494–9.PubMed CentralView ArticlePubMedGoogle Scholar
  76. Ballatory N, Krance SM, Notenboom S, Shi S, Tieu K, Hammond CL. Glutathione dysregulation and the etiology and progression of human disease. Biol Chem. 2009;390(3):191–214.Google Scholar
  77. Kharb S. Low whole blood glutathione levels in pregnancies complicated by preeclampsia and diabetes. Clin Chim Acta. 2000;294:179–83.View ArticlePubMedGoogle Scholar
  78. Boden G, Duan X, Homko C, Molina EJ, Song W, Perez O, et al. Increase in endoplasmic reticulum stress-related proteins and gene in adipose tissue of obese, insulin-resistant individuals. Diabetes. 2008;57:2438–44.PubMed CentralView ArticlePubMedGoogle Scholar
  79. Fuentes-Almagro CA, Prieto-Álamo M-J, Pueyo C, Jurado J. Identification of proteins containing redox-sensitive thiols after PRDX1, PRDX3 and GCLC silencing and/or glucose oxidase treatment in Hepa 1–6 cells. J Prot. 2012;77:262–79.View ArticleGoogle Scholar
  80. Nonn L, Berggren M, Powis G. Increased expression of mitochondrial peroxiredoxin-3 (thioredoxin peroxidase-2) protects cancer cells against hypoxia and drug-induced hydrogen peroxide-dependent apoptosis. Mol Cancer Res. 2003;1:682–9.PubMedGoogle Scholar
  81. Chang T-S, Cho C-S, Park S, Yu S, Kang SW, Rhee SG. Peroxiredoxin III, a mitochondrion-specific peroxidase, regulates apoptotic signaling by mitochondria. J Biol Chemis. 2004;279(40):41975–84.View ArticleGoogle Scholar
  82. Janciauskiene S, Larsson S, Larsson P, Virtala R, Jansson L, Stevens T. Inhibition of lipopolysaccharide-mediated human monocyte activation, in vitro, by α1-antitrypsin. Biochem Biophys Res Comm. 2004;321:592–600.View ArticlePubMedGoogle Scholar
  83. Stockley RA, Shaw J, Afford SC, Morrison HM, Burnett D. Effect of alpha-1-proteinase inhibitor on neutrophil chemotaxis. Am J Respir Cell Mol Biol. 1990;2(2):163–70.View ArticlePubMedGoogle Scholar
  84. Bergin DA, Reeves EP, Meleady P, Henry M, McElvaney OJ, Carroll TP, et al. α-1 Antitrypsin regulates human neutrophil chemotaxis induced by soluble immune complexes and IL-8. J Clin Invest. 2010;120(12):4236–50.PubMed CentralView ArticlePubMedGoogle Scholar
  85. Al-Omari M, Korenbaum E, Ballmaier M, Lehmann U, Jonigk D, Manstein DJ, et al. Acute-phase protein α1-antitrypsin inhibits neutrophil calpain I and induces random migration. Mol Med. 2011;17:865–74.PubMed CentralView ArticlePubMedGoogle Scholar
  86. Griese M, Latzin P, Kappler M, Weckerle K, Heinzlmaier T, Bernhardt T, et al. α1-antitripsin inhalation reduces airway inflammation in cystic fibrosis patients. Eur Respir J. 2007;29:240–50.View ArticlePubMedGoogle Scholar
  87. Kalis M, Kumar R, Janciauskiene S, Salehi A, Cílio CM. α1-antitripsinenhances insulin secretion and prevents cytokine-mediated apoptosis in pancreatic β-cells. Islet. 2010;2(3):185–9.View ArticleGoogle Scholar
  88. Lu Y, Tang M, Wasserfall C, Kou Z, Campbell-Thompson M, Gardemann T, et al. α1-antitrypsin gene therapy modulates cellular immunity and efficiently prevents type 1 diabetes in nonobese diabetic mice. Human Gene Terapy. 2006;17:625–34.View ArticleGoogle Scholar
  89. Oliva K, Barker G, Rice GE, Bailey MJ, Lappas M. 2D-DIGE to identify proteins associated with gestational diabetes in omental adipose tissue. J Endocrinol. 2013;218:165–78.View ArticlePubMedGoogle Scholar
  90. Laemmli UK. Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature. 1970;227(5259):680–5.View ArticlePubMedGoogle Scholar

Copyright

© de Souza et al. 2015

Advertisement