Prenatal stress mediates tissue specific programming of 11β-hydroxysteroid dehydrogenase type 1 expression in diet-induced obese and dietary resistant rats

doi:10.21203/rs.3.rs-4650919/v1

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Prenatal stress mediates tissue specific programming of 11β-hydroxysteroid dehydrogenase type 1 expression in diet-induced obese and dietary resistant rats

https://doi.org/10.21203/rs.3.rs-4650919/v1

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Background

The link between uterine stress and fetal programming of metabolic diseases in the offspring is well established. However, susceptibility to prenatal stress-induced programming of metabolic diseases is not uniform across population. We have previously reported that diet-induced obese rats (DIO) are more susceptible to develop metabolic diseases in response to prenatal stress and postnatal HF diet challenge when compared to dietary resistant rats (DR), but the mechanism behind this phenomenon is unclear. In the present study, we hypothesized that prenatally stressed DIO rats will have alterations in stress axis and 11βHSD1 expression, which in turn predisposes them to metabolic disorders.

Methods

Pregnant DIO and DR rats were divided into 2 groups: control (normal pregnancy) and prenatal stress (restraint stress during the last week of gestation) groups. The male offspring were weaned onto chow diet until 9 weeks of age and then shifted to either chow or HF diet (45% calories from fat) for 1 week. Following 1 week of chow/HF diet treatment, the animals were sacrificed, and tissues were collected for assessing stress axis activity and 11βHSD1 expression.

Results

Prenatal stress activated stress axis with increased NE levels in the PVN and CRH levels in the median eminence in both DIO and DR rats. Though there was no change in either circulating CORT levels or adrenal gland weight, 11βHSD1 protein expression increased significantly in adipose tissue of prenatally stressed DIO and DR offspring compared to controls. Baseline levels of 11βHSD1 protein in the adipose tissue were higher in DIO than DR animals implicating a role for 11βHSD1 in the development of obesity. Postnatal HF diet did not have any additive effect on prenatal stress-induced changes.

Conclusion

Our results suggest that stress axis activation with induction of adipose tissue 11βHSD1 might be involved in the fetal origins of metabolic diseases.

Health sciences/Endocrinology/Endocrine system and metabolic diseases/Obesity

Biological sciences/Physiology/Metabolism/Metabolic diseases/Obesity

11β-hydroxysteroid dehydrogenase type 1

obesity

adipose tissue

corticosterone

diet-induced obese and dietary resistant

Growing evidence from animal and human epidemiological studies supports the idea that prenatal stress is not only associated with increased susceptibility to develop psychological disorders but also metabolic disorders like obesity and diabetes^1,2. Using diet-induced obese (DIO) and dietary resistant (DR) animal model, we have previously demonstrated that prenatal stress combined with postnatal HF feeding predisposes the offspring to adverse metabolic outcomes like hyperinsulinemia³. In addition, our results also suggest that individuals with pre-existing genetic predisposition to obesity (diet-induced obese, DIO rats) are more susceptible to the programming effects of prenatal stress when compared with their counterparts (dietary resistant, DR rats)³. Though the association between prenatal stress and metabolic disorders is well established⁴, the mechanism behind this phenomenon is far from clear.

The established association of Cushing’s syndrome with excess circulating corticosterone and metabolic syndrome opens up the possibility that dysregulation of stress axis function could be one of the mechanisms through which prenatal stress programs the offspring for metabolic diseases⁵. Stress axis activation commences with the activation of corticotropin releasing hormone (CRH) neurons in the paraventricular nucleus (PVN) of the hypothalamus. Though a variety of neurotransmitters like 5-HT and GABA play an important role in modulating CRH neuronal activation, norepinephrine (NE) from the brain stem nuclei provides the primary stimulus for CRH synthesis and release⁶. Upon activation, CRH is released in the median eminence and then transported to the pituitary through the hypothalamo-hypophyseal portal system. In the pituitary, CRH acts on the corticotrophs to release adrenocorticotropin (ACTH) which then enters the systemic circulation and acts on the adrenal cortex to release glucocorticoids (cortisol in humans and corticosterone (CORT) in rodents)⁷. The actions of CORT in the peripheral tissues are not only dependent on its circulating levels, but also on the tissue levels of glucocorticoid receptors and the activity of the enzyme, 11-β hydroxysteroid dehydrogenase type 1 (11βHSD1)^8,9. 11βHSD1 is a NADPH dependent microsomal enzyme that is highly expressed in metabolically active tissues like liver, adipose tissue and also in the central nervous system^10–12. In vivo, 11βHSD1 acts as a reductase and converts inactive 11-dehydrocorticosterone to CORT in rodents or inactive cortisone to cortisol in humans^9,13. Despite low circulating CORT, the intracellular levels of CORT available for binding with GR may be increased by increased 11βHSD1 in the target tissues. Hence, increased 11βHSD1 expression (either independently or with concurrent hyperactivation of stress axis) results in tissue-specific amplification of glucocorticoid actions and implicated in the pathogenesis of obesity and insulin resistance^14–16.

Previous studies using prenatal stress programming model have demonstrated higher corticosterone secretion at the end of the lighting period in both males and female offspring¹⁷, increase in CRH expression in the PVN¹⁸ and enhanced responsiveness to stressors in the postnatal period resulting in higher circulating ACTH and cortisol levels¹⁹. Further, dexamethasone injection during late gestation resulted in persistent elevations in 11βHSD1 expression and activity in the liver and adipose tissue²⁰. Similarly, maternal nutrient restriction during pregnancy and gestational diabetes also increased the expression of GR and 11βHSD1 in the adipose tissue^21,22. However, it is not clear if stress axis dysregulation and/or increased 11βHSD1 expression in the peripheral tissues could be the underlying mechanism behind the metabolic effects induced by prenatal stress. The overall aim of the present study is to test the hypothesis that prenatally stressed DIO animals will have hyperactivation of stress axis and/or increased 11βHSD1 in liver and adipose tissue, which in turn leads to metabolic disorders.

Animals and Treatment

Adult female and male DIO and DR rats (8 weeks old) were obtained from Charles River Laboratories (Wilmington, MA). All the animals were housed in temperature-controlled room (23 ± 2ºC) in 12:12 hours light dark cycle with ad libitum feed and water. For the experiment, female DIO (n = 8) and DR rats (n = 9) were housed individually with their male counterparts. Pregnancy was confirmed by spotting sperms in stained vaginal smears obtained from the female DIO and DR rats. Once confirmed, pregnant DIO and DR rats were classified into 2 groups: non-stressed controls (underwent normal pregnancy without any disturbance) and prenatal stress group (underwent restraint stress in a transparent glass cylinder from gestational day 14–21 for 45 minutes thrice a day). We used a variable time restraint stress protocol to prevent the animals from getting accustomed to the time of restraint stress²³. Once the pups were born, the litter size was normalized to 8 pups/litter (4 males and 4 females) to exclude the effects of litter size on the nutritional status of the offspring. Male offspring were weaned onto chow diet until 9 weeks of age. After that, the DIO and DR male offspring were divided into 2 groups each (n = 6–10/diet group, 2 from each mom) and shifted to either chow diet (23% protein, 72% carbohydrate, and 5% calories as fat with an energy density of 3.11 kcal/g) or high fat diet (20% protein, 35% carbohydrate, and 45% calories as fat with an energy density of 4.73 kcal/g; Research Diets, New Brunswick, NJ). Following 1 week of chow or HF diet treatment, the animals were sacrificed by decapitation around noon time and trunk blood, liver, visceral adipose tissue, and brain tissue were collected. Adrenal glands were also collected and weighed. Serum was separated from trunk blood after centrifugation and stored at -80ºC for CORT measurements. All procedures involving animals were in compliance with NIH’s guide for the care and use of laboratory animals and were approved by the IACUC at Michigan State University.

Brain microdissection

Serial brain sections of 300µm thickness were obtained using a cryostat (Slee Mainz, London) maintained at -20°C and then transferred to a cold stage maintained at the same temperature. The Paraventricular nucleus (PVN) and median eminence (ME) were micro dissected using the Palkovits’ microdissection technique as described previously using a 500µm punch. Nuclei were identified with the help of a rat brain atlas²⁴. Care was taken to include all subdivisions of each nucleus. Tissues were stored at -80°C until analysis.

HPLC-EC

NE in PVN punches was measured using HPLC-EC as described previously²⁵. Briefly, the HPLC-EC system consisted of a LC 20-AD pump (Shimadzu, Columbia, MD), and a phase II, 5-µm ODS reverse phase C-18 column (Phenomenex, Torrance, CA), a SIL-20AC autoinjector, a CTO-20AC column oven maintained at 37°C and a LC-4C detector (Bioanalytical Systems, West Lafayette, IN). The mobile phase contained chloroacetic acid (14.5 g/L), octane sulfonic acid (0.3 g/L), EDTA (0.25 g/L) and sodium hydroxide (4.675 g/L). The pH was adjusted to 3.1 and the mobile phase was filtered and degassed. Acetonitrile 17.5 ml and 13 ml of tetrahydrofuran were added to the mobile phase. The flow rate of the mobile phase was 1.8 ml/min. The sensitivity of the detector was 1 nA full scale, and the potential of the working electrode was 0.65 V. Micro dissected hypothalamic nuclei were homogenized in 50 µl of 0.1M perchloric acid and 5–10 µl of the homogenate was used for protein estimation. The rest of the homogenate was centrifuged at 5000rpm for 5 minutes and 15 µl of the supernatant was loaded with 15 µl of the internal standard (dihydroxybenzylamine, 0.05 M) in the autoinjector and injected into the HPLC system. Chromatograms were analyzed for NE concentrations using the Class VP software v. 7.2 (Shimadzu, Columbia, MD).

Western blotting

Liver tissue was homogenized in lysis buffer (20mM Tris, 150mM Sodium Chloride, 1mM EDTA, 10% Glycerol and 1% Triton-X) with addition of protease inhibitors (0.5µl of 100mM PMSF, 0.1µl of 100mM Sodium Orthovanadate and Aprotinin). The homogenates were centrifuged at 13,000 rpm for 20 minutes at 40C. Protein levels in the supernatant were estimated using Micro BCA protein assay (Pierce Biochemicals, Rockford, IL). Equal amounts of protein (80µg) were resolved on 4–20% NuSep Precast gels (NuSep Inc, Bogart, GA) and transferred to a nitrocellulose membrane. The membranes were blocked for 1 hour at room temperature using Odyssey Blocking buffer (Licor Biosciences, Lincoln, NE). The membranes were then incubated with the following primary antibodies: 1:500 anti-11βHSD1 rabbit polyclonal IgG (Santa Cruz Biotechnology Inc, Santa Cruz, CA) and 1:1000 anti- β actin goat polyclonal IgG (for liver tissues) (Abcam, Cambridge, MA) diluted in odyssey blocking buffer with 0.1% tween-20 and 1% thiomersal and incubated overnight at 4°C. The membranes were washed 4 times with PBS + 0.1% Tween 20 for 5 minutes each. After washing, the membranes were incubated with infra-red dye conjugated secondary antibodies (donkey anti-rabbit IgG (1:15,000) and donkey anti-goat IgG (1:20,000) (Licor Biosciences, Lincoln, NE) for 50 minutes at room temperature. The membranes were washed again with PBS + 0.1% Tween 20 for 4 times. The immunoreactive bands were visualized in a Laser scanner (Licor Biosciences, Lincoln, NE) and the intensity of the bands was measured using Image J software (NIH, Bethesda, MD). The values were normalized to the loading controls (β-actin) and expressed as a ratio of 11βHSD1/β-actin.

Enzyme-Linked Immunosorbent Assay (ELISA)

CRH levels in the median eminence were measured using an EIA kit (Phoenix Pharmaceuticals, Burlingame, CA). Briefly, tissue punches of the median eminence were homogenized in 125µl of Phosphate buffered saline (PBS) using an ultrasound sonicator (Kontes, Vineland, NJ). From the homogenate, 50µl was used in duplicates for the measurement of CRH using EIA and 10µl was used for protein estimation. The CRH values were expressed as ng/µg of protein.

Quantitative RT-PCR

Total RNA was extracted from the liver and adipose tissue (n = 5–8/group) using GenElute Mammalian Total RNA Miniprep kit (Sigma Aldrich, St. Louis, MO, USA) and RNeasy Lipid Mini kit (Qiagen, Valencia, CA) respectively. RNA quality was determined using Nanodrop Spectrophotometer (Thermo Scientific, Wilmington, DE, USA) and the samples with low quality RNA assessed by 260/280 ratio (< 1.8 or > 2.1) were excluded from further analysis. Equal quantities of RNA (500ng) from the samples were subjected to reverse transcription for cDNA conversion with High-Capacity cDNA reverse transcription kit (Applied Biosystems, Carlsbad, CA). PCR reactions were carried out on ABI 7500 Fast real time PCR system (Applied Biosystems, Carlsbad, CA) using SYBR Green master mix (SA Biosciences, Frederick, MD). The PCR reaction mixture included 1 µl cDNA (25ng), 10.5 µl RNA grade water, 1 µl primer and 12.5 µl SYBR Green master mix. The PCR conditions were the following: holding stage (95⁰C for 10 minutes) and cycling stage (95⁰C for 15 sec, 60⁰C for 1 minute and 72⁰C for 35 seconds) followed by melt curve to confirm the specificity of the amplified products. The primers for 11βHSD1 (Ref. Seq. No. NM_017080) and β-actin (Ref. Seq. No. NM_031144) were purchased from SA Biosciences, Frederick, MD. The Ct values were normalized to the house keeping gene, β-actin. The fold change was calculated by 2^∆∆CT method.

Immunohistochemistry

Formalin fixed specimens were processed, embedded in paraffin, and sectioned on a rotary microtome. Sections (4 µm) were placed on adhesive slides and dried at 56°C overnight. The slides were subsequently deparaffinized in Xylene and hydrated through descending grades of ethanol to distilled water. Slides were placed in Tris Buffered Saline (pH 7.5) for 5 minutes for pH adjustment. Heat was Induced in a rice steamer for 30 minutes (followed by 10 minutes at room temperature) followed by subsequent rinses and blocking for endogenous peroxidase using 3% Hydrogen Peroxide/Methanol bath (1:4 ratio) for 30 minutes at room temperature followed by rinsing with running tap and distilled water. Slides are blocked with normal goat serum (Vector Labs, Burlingame, CA) for 30 minutes. Endogenous Biotin was blocked by incubation with Avidin D (Vector) and d-Biotin (Sigma Aldrich, St.Louis, MO) for 15 minutes in each reagent. The slides were then incubated with rabbit Polyclonal anti-11β-HSD1 antibody (Santa Cruz, CA) diluted in Normal Antibody Diluent (NAD, 1:100) (Scytek, Logan, UT) overnight at 4°C. After washing, the slides were incubated with biotinylated goat anti-rabbit and IgG H + L (Vector) (11ug/ml) for 30 minutes followed by treatment with VectaStain Elite ABC Reagent (Vector) for 30 minutes. Reaction was developed with Nova Red (Vector) for 15 minutes, followed by counterstain with Gill 2 Hematoxylin (Richard Allen, Kalamazoo, MI) for 10 seconds, differentiated in 1% aqueous glacial acetic acid and rinsed in running tap water. Slides were then dehydrated through ascending grades of ethanol; cleared through several changes of Xylene and cover-slipped using Flotex (Lerner, Pittsburgh PA) permanent mounting media.

Statistics:

Changes in stress axis parameters and 11βHSD1 expression were analyzed by ANOVA followed by Fishers LSD test. All the statistical analyses were performed in Statview software. Three-way ANOVA analysis was performed to assess the interaction between stress, genotype, and diet.

Effects of prenatal stress and postnatal HF diet on stress axis in the DIO and DR offspring

NE levels in the paraventricular nucleus

Changes in NE levels (mean ± SE, pg/µg of protein) in the PVN of DIO and DR offspring subjected to prenatal stress and 1-week postnatal HF diet is shown in Fig. 1A. Prenatal stress significantly increased NE levels in the DR animals (p < 0.05) when compared with their controls, irrespective of diet treatment. In DIO animals, prenatal stress by itself did not cause an increase in PVN NE levels. However, 1 week HF diet treatment greatly increased PVN NE levels in the stressed DIO offspring (p < 0.05). The increase in NE levels observed in stressed DIO offspring on HF diet was significantly higher compared to all other DIO and DR groups. We observed a significant stress effect (F = 23.6, p < 0.0001), genotype effect (F = 7.1, p < 0.05) and a trend towards significant stress x genotype x diet effect (F = 3.37, p = 0.07).

CRH levels in the median eminence

CRH levels (mean ± SE, pg/µg of protein) in the ME of DIO and DR rats after prenatal stress and 1-week postnatal HF diet is shown in Fig. 1B. In the DR animals, CRH levels in the ME were significantly higher in the stressed group (226.07 ± 32.9 and 240.7 ± 21.4 in chow and HF groups respectively) compared to the non-stressed controls (70.5 ± 8.9 and 83.2 ± 12.1 in chow and HF fed groups), irrespective of the diet treatment. Similarly, prenatally stressed DIO animals (168.1 ± 26.2 and 208.6 ± 29.7 in chow and HF groups respectively, p < 0.05) had significantly higher CRH levels in the ME compared to the non-stressed DIO animals (73.1 ± 14.9 and 95.4 ± 7.5 in chow and HF fed groups respectively). We observed an overall effect of stress (F = 78.7, p < 0.0001) and diet (F = 5.3, p < 0.05) on increasing CRH levels in the ME in DIO and DR animals.

Serum Corticosterone

Serum Corticosterone (mean ± SE, ng/ml) levels measured at the time of sacrifice after 1 week chow/HF diet exposure in prenatally stressed DIO and DR offspring is shown in Fig. 1C. Unlike NE and CRH levels, prenatal stress did not alter serum corticosterone levels in both DIO and DR animals. HF diet exposure for 1 week also did not have any effect on corticosterone levels in both DIO and DR animals.

Adrenal gland weight to body weight ratio

Adrenal gland to BW ratio (mean ± SE) in prenatally stressed DIO and DR male offspring after exposure to 1 week chow or HF diet is shown separately in Fig. 1D. In correlation with serum corticosterone levels, there were no significant differences in the adrenal gland to body weight ratio between non-stressed and stressed DIO and DR groups. However, the adrenal gland/BW ratio was significantly lower in the DIO groups when compared with the DR groups (p < 0.05). We observed a significant genotype effect (F = 44.1, P < 0.0001) with respect to adrenal gland/BW ratio.

Effects of prenatal stress and postnatal HF diet on 11βHSD1 expression

Hepatic 11βHSD1 expression

Hepatic 11βHSD1 mRNA and protein levels in the DIO and DR groups were shown in Fig. 2A and B respectively. 11βHSD1 mRNA levels were expressed as 11βHSD1 mRNA: β-actin ratio. Prenatal stress did not have any effect on 11βHSD1 mRNA expression in the liver in the DIO and DR groups. However, the fold change in 11βHSD1 mRNA levels in the prenatally stressed DIO offspring (chow and HF diet) was significantly (p < 0.05) higher than their corresponding DR groups. HF diet exposure for 1 week produced a significant (p < 0.05) down-regulation in 11βHSD1 mRNA expression in the liver in all the DIO and DR groups compared to their respective chow-fed counterparts. We observed a significant diet (F = 50.05, P < 0.0001) and genotype (F = 12.15, p < 0.005) but no prenatal stress effect in 11βHSD1 mRNA expression in the liver in both the DIO and DR groups. Similarly, the protein levels of 11βHSD1 measured by western blotting also showed no differences between non-stressed and stressed DIO and DR groups.

Adipose tissue 11βHSD1 expression

11βHSD1 mRNA levels in the visceral adipose tissue were expressed as 11βHSD1: β-actin ratio (Fig. 3A). On chow diet, prenatal stress produced a moderate increase (1.3 ± 0.1-fold change, p < 0.05) in the mRNA expression of 11βHSD1 in the VAT of DIO offspring when compared to its non-stressed counterparts (0.99 ± 0.02) and stressed DR chow (0.90 ± 0.02) and HF animals (0.80 ± 0.12). Exposure to 1 week HF diet produced a significant down-regulation in the mRNA expression of 11βHSD1 in the chronic stress DIO offspring. In the DR offspring, prenatal stress did not produce any changes in the 11βHSD1 expression in the VAT. We observed a significant diet (F = 9.9, P < 0.005) and stress x genotype interaction (F = 4.3, p < 0.05) in 11βHSD1 mRNA expression in the liver in both the DIO and DR groups.

11βHSD1 protein levels in the VAT were expressed as the percentage of adipocytes positive for 11βHSD1 staining per field in the DIO and DR offspring and depicted in Fig. 3B. Representative images for 11βHSD1 staining in the VAT are shown in Fig. 4. In both the DIO and DR offspring, irrespective of the diet, stressed offspring had higher percentage of adipocytes that expressed 11βHSD1 protein than their respective non-stressed controls (both chow and HF groups). Although, there was no difference between stressed DIO and DR offspring, interestingly the baseline expression of 11βHSD1 protein in the VAT of DIO controls (20.0 ± 1.5, p < 0.05) were significantly higher than the DR controls (4.7 ± 0.5) confirming the role of VAT 11βHSD1 in the development of obesity. We observed a significant stress (F = 131.5, p < 0.0001), diet (F = 8.5, p < 0.05) and stress x diet x genotype interaction (F = 7.7, p < 0.05) with respect to VAT 11βHSD1 protein expression.

Extending our previous findings, we report based on the results from the present study that i) Prenatal stress increases stress axis activity as marked by increases in NE levels in the PVN and CRH levels in the median eminence both in DR and DIO animals, irrespective of postnatal HF diet treatment ii) This central hyperactivation of stress axis was not accompanied by increases in circulating CORT levels or adrenal gland weight iii) Prenatal stress resulted in up regulation of 11βHSD1 specifically in the adipose tissue but not in the liver suggesting that prenatal stress induces tissue specific programming of 11βHSD1 expression both in DR and DIO animals.

Our results are in agreement with other studies examining the effects of prenatal stress on stress axis function in terms of NE and CRH concentrations in the offspring. Prenatal stress has been reported to increase NE concentrations in the hypothalamus²⁶ and increase NE turnover rate in the brain stem suggesting higher NE release²⁷. Also, CRH concentrations in the amygdala, which is another stress regulating center in the brain, was increased in the prenatally stressed offspring²⁸. Apart from being a part of stress axis, PVN also regulates feeding behavior. Acute and chronic infusion of NE in the PVN produces hyperphagia and promotes obesity^29,30. Hence, the increase in PVN NE concentrations observed in the prenatally stressed offspring might lead to hyperphagia in the offspring’s adulthood.

In the current study, despite increases in NE and CRH levels, we did not observe any changes in baseline CORT levels between stressed and non-stressed DIO and DR offspring. Previously, studies have demonstrated a similar finding where there was no difference in baseline CORT levels in prenatally stressed offspring at postnatal day (PND) 21 ^2,31 and PND90³¹. On the other hand, few other studies have also showed an increase in baseline CORT in the offspring subjected to prenatal stress³² and prolonged CORT response to stress³³. The disparity in the findings might be due to the differences in the stress protocol and the age at which CORT measurements were made in the offspring. Further, based on the results from the present study, it is still not clear whether increased renal clearance have masked the true CORT levels in the prenatally stressed DIO/DR offspring. Additional studies on CORT metabolite measurements in the urine are needed to answer this question.

Despite no detectable changes in circulating CORT levels, glucocorticoids might still play a role in the peripheral metabolic outcomes observed in the prenatally stressed DIO offspring through the enzyme, 11βHSD1. 11βHSD1 is involved in pre-receptor metabolism of CORT in the metabolically active peripheral tissues like liver and the adipose tissue where it converts inactive 11-dehydroCORT to active CORT^8,9. Irrespective of circulating CORT levels, increase in 11βHSD1 expression in these tissues will result in amplification of glucocorticoid actions leading to obesity and insulin resistance⁸. In the present study, prenatal stress mediated increase in 11βHSD1 protein expression was tissue specific as we observed it specifically in the adipose tissue but not in the liver. Supporting tissue-specific programming of 11βHSD1 expression, studies have shown that dexamethasone treatment during late gestation and diabetic pregnancy results in offspring with higher 11βHSD1 expression in the liver and adipose tissue^20,21. Similarly, postnatal overfeeding, also known to program the offspring for obesity, results in higher 11βHSD1 expression in the adipose tissue³⁴. Interestingly, prenatal stress also resulted in increased 11βHSD1 protein expression in the DR offspring, which did not have any adverse metabolic outcomes and stress axis hyperactivation as observed in DIO animals. Whether this has any physiological significance in the DR animals will remain a question until further experiments on chronic HF diet exposure are carried out. Also, prenatally stressed offspring with elevated 11βHSD1 expression in the adipose tissue are not overtly obese when compared with their non-stressed counterparts. In concordance with our results, maternal perinatal undernutrition and dexamethasone administration during late gestation also resulted in the offspring with a lean phenotype but with a lipogenic adipose tissue gene profile^20,35. Taken together, these studies suggest that elevated 11βHSD1 expression precedes obesity in by fetal programming.

There are several mechanisms by which increased 11βHSD1 and its resultant increase in intra-adipocyte CORT could contribute to the development of obesity and metabolic syndrome. In the adipose tissue, glucocorticoids regulate the activity and expression of lipoprotein lipase (LPL)³⁶, an enzyme expressed in the endothelial cells that catalyzes the conversion of circulating triglycerides to FFA which is then re-esterified to be stored in the adipocytes³⁷. Higher LPL activity results in more triglyceride accumulation and hypertrophy of the adipocytes leading to an unhealthy pro-inflammatory and insulin resistant adipose tissue. This condition is further compounded by the anti-lipolytic actions of insulin during the hyper insulinemic state³⁸. Glucocorticoids also promote pre-adipocyte differentiation and proliferation resulting in increase in fat mass. In addition to lipogenesis, glucocorticoids can also cause lipolysis by stimulating hormone sensitive lipase in the adipose tissue which increases FFA flux into the portal circulation and thus facilitates ectopic fat distribution in liver and muscle³⁹.

In summary, the present study suggests that prenatal stress results in hyperactivation of stress axis with simultaneous up regulation of 11βHSD1 in the visceral adipose tissue of the offspring. Although, both DIO and DR offspring had increased 11βHSD1 in the adipose tissue, DR offspring exhibited a healthy metabolic phenotype. Hence, 11βHSD1 could mediate the metabolic effects observed in the prenatally stressed offspring and thus may be a potential mechanism for fetal origins of obesity.

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Prenatal stress mediates tissue specific programming of 11β-hydroxysteroid dehydrogenase type 1 expression in diet-induced obese and dietary resistant rats

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Version 1

Abstract

Figures

Introduction

Methods

Animals and Treatment

Brain microdissection

HPLC-EC

Western blotting

Enzyme-Linked Immunosorbent Assay (ELISA)

Quantitative RT-PCR

Immunohistochemistry

Statistics:

Results

NE levels in the paraventricular nucleus

CRH levels in the median eminence

Serum Corticosterone

Adrenal gland weight to body weight ratio

Effects of prenatal stress and postnatal HF diet on 11βHSD1 expression

Hepatic 11βHSD1 expression

Adipose tissue 11βHSD1 expression

Discussion

References

Additional Declarations

Status:

Version 1