Effect of insulin on AKR1C3 expression in female adipose tissue: in-vivo and in-vitro study of adipose androgen generation in polycystic ovary syndrome

Abstract Objectives Studies suggest that inflammation mediates the link between obesity and its comorbidities including type 2 diabetes and cardiovascular disease. Hence, there is a demand for effective alternative or complementary approaches to treat obesity-associated inflammation. The objective of this study was to determine whether consumption of blackberries (BL) and raspberries (RB) alone or in combination reduce obesity-induced inflammation. Methods In Vitro Study: RAW 264.7 macrophages were pretreated with either BL, RB, or BL + RB, each at a final concentration of 200 µg/mL for 2 h. LPS (1 ng/mL) was then added to the media for 16 h. mRNA expression of inflammatory cytokines was measured. In Vivo Study: Five-week-old mice were acclimated to a low-fat low-sucrose (LFLS) diet for one week after which mice were randomized 10 per group to one of five groups: 1) LFLS, 2) high-fat high-sucrose (HFHS), 3) HFHS + 10% BL, 4) HFHS + 10% RB, or 5) HFHS + 5% BL + 5% RB. Expression of inflammatory markers was measured in the liver as well as epididymal and inguinal white adipose tissue. Results In Vitro Study: Each berry alone and in combination suppressed the LPS-induced increase in inflammatory markers, with the combination (BL + RB) having the greatest effect. The combination suppressed LPS-induced expression of Ccl2, Tnfa, F4/80, and Il6 by 3.7−, 5.3−, 5.3−, and 4.4-fold, respectively. In Vivo Study: Gene expression analysis indicated that berry consumption had no significant effect on proinflammatory (Ccl2, Il1b, Tnfa, Il6, Itgam) or anti-inflammatory (Adipoq, Arg1, Mgl1) markers in adipose tissue depots or liver. However, relatively low gene expression of inflammatory markers in the tissues indicates that the mice fed the HFHS diet failed to develop a robust inflammatory state. Conclusions BL and RB have direct anti-inflammatory effects on immune cells. Initial analysis indicates that consumption of BL and RB has no significant effects on markers of inflammation in a diet-induced mouse model of obesity. However, it is possible that the relatively low levels of inflammation in these mice masked the anti-inflammatory potential of BL and RB. Ongoing analysis will provide additional insights into the effects of BL and RB on inflammation in these tissues. Funding Sources Lewis Foundation Award.

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Regulation of Androgen Receptor Expression Alters AMPK Phosphorylation in the Endometrium: In Vivo and In Vitro Studies in Women with Polycystic Ovary Syndrome

International Journal of Biological Sciences ◽

10.7150/ijbs.13109 ◽

2015 ◽

Vol 11 (12) ◽

pp. 1376-1389 ◽

Cited By ~ 17

Author(s):

Xin Li ◽

Bano Pishdari ◽

Peng Cui ◽

Min Hu ◽

Hong-Ping Yang ◽

...

Keyword(s):

Androgen Receptor ◽

Polycystic Ovary Syndrome ◽

Polycystic Ovary ◽

Receptor Expression ◽

In Vitro Studies ◽

Androgen Receptor Expression ◽

Ovary Syndrome ◽

Ampk Phosphorylation

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PAPSS2 Deficiency Causes Androgen Excess via Impaired DHEA Sulfation—In Vitro and in Vivo Studies in a Family Harboring Two Novel PAPSS2 Mutations

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2014-3556 ◽

2015 ◽

Vol 100 (4) ◽

pp. E672-E680 ◽

Cited By ~ 38

Author(s):

Wilma Oostdijk ◽

Jan Idkowiak ◽

Jonathan W. Mueller ◽

Philip J. House ◽

Angela E. Taylor ◽

...

Keyword(s):

Polycystic Ovary Syndrome ◽

Polycystic Ovary ◽

Challenge Test ◽

Compound Heterozygous ◽

Androgen Excess ◽

Ovary Syndrome ◽

Functional Impact ◽

Low Serum

Context: PAPSS2 (PAPS synthase 2) provides the universal sulfate donor PAPS (3′-phospho-adenosine-5′-phosphosulfate) to all human sulfotransferases, including SULT2A1, responsible for sulfation of the crucial androgen precursor dehydroepiandrosterone (DHEA). Impaired DHEA sulfation is thought to increase the conversion of DHEA toward active androgens, a proposition supported by the previous report of a girl with inactivating PAPSS2 mutations who presented with low serum DHEA sulfate and androgen excess, clinically manifesting with premature pubarche and early-onset polycystic ovary syndrome. Patients and Methods: We investigated a family harboring two novel PAPSS2 mutations, including two compound heterozygous brothers presenting with disproportionate short stature, low serum DHEA sulfate, but normal serum androgens. Patients and parents underwent a DHEA challenge test comprising frequent blood sampling and urine collection before and after 100 mg DHEA orally, with subsequent analysis of DHEA sulfation and androgen metabolism by mass spectrometry. The functional impact of the mutations was investigated in silico and in vitro. Results: We identified a novel PAPSS2 frameshift mutation, c.1371del, p.W462Cfs*3, resulting in complete disruption, and a novel missense mutation, c.809G>A, p.G270D, causing partial disruption of DHEA sulfation. Both patients and their mother, who was heterozygous for p.W462Cfs*3, showed increased 5α-reductase activity at baseline and significantly increased production of active androgens after DHEA intake. The mother had a history of oligomenorrhea and chronic anovulation that required clomiphene for ovulation induction. Conclusions: We provide direct in vivo evidence for the significant functional impact of mutant PAPSS2 on DHEA sulfation and androgen activation. Heterozygosity for PAPSS2 mutations can be associated with a phenotype resembling polycystic ovary syndrome.

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In vivo and in vitro exposure to free fatty acids induce increased androgen production in animal models: a potential pathogenic factor of the polycystic ovary syndrome

Fertility and Sterility ◽

10.1016/j.fertnstert.2008.07.388 ◽

2008 ◽

Vol 90 ◽

pp. S16

Author(s):

C.G. Baptiste ◽

X. Ci ◽

L. Chouinard ◽

N. Gallot-Payet ◽

A.C. Carpentier ◽

...

Keyword(s):

Fatty Acids ◽

Animal Models ◽

Polycystic Ovary Syndrome ◽

Free Fatty Acids ◽

Polycystic Ovary ◽

Pathogenic Factor ◽

Ovary Syndrome ◽

In Vitro Exposure

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Enhanced Thecal Androgen Production Is Prenatally Programmed in an Ovine Model of Polycystic Ovary Syndrome

Endocrinology ◽

10.1210/en.2011-1607 ◽

2012 ◽

Vol 153 (1) ◽

pp. 450-461 ◽

Cited By ~ 44

Author(s):

Kirsten Hogg ◽

Julia M. Young ◽

Elizabeth M. Oliver ◽

Carlos J. Souza ◽

Alan S. McNeilly ◽

...

Keyword(s):

Polycystic Ovary Syndrome ◽

Polycystic Ovary ◽

Female Offspring ◽

Fetal Life ◽

Ovary Syndrome ◽

Androgen Synthesis ◽

Basal Expression ◽

Thecal Cells

One of the hallmarks of polycystic ovary syndrome (PCOS) is increased ovarian androgen secretion that contributes to the ovarian, hormonal, and metabolic features of this condition. Thecal cells from women with PCOS have an enhanced capacity for androgen synthesis. To investigate whether this propensity is a potential cause, rather than a consequence, of PCOS, we used an ovine prenatal androgenization model of PCOS and assessed ewes at 11 months of age. Pregnant Scottish Greyface ewes were administered 100 mg testosterone propionate (TP) or vehicle control twice weekly from d 62 to 102 of gestation, and female offspring (TP = 9, control = 5) were studied. Prenatal TP exposure did not alter ovarian morphology or cyclicity, or plasma androgen, estrogen, and gonadotropin concentrations, at this stage. However, follicle function was reprogrammed in vivo with increased proportions of estrogenic follicles (P < 0.05) in the TP-exposed cohort. Furthermore, in vitro the thecal cells of follicles (>4 mm) secreted more LH-stimulated androstenedione after prenatal androgenization (P < 0.05), associated with increased basal expression of thecal StAR (P < 0.01), CYP11A (P < 0.05), HSD3B1 (P < 0.01), CYP17 (P < 0.05), and LHR (P < 0.05). This provides the first evidence of increased thecal androgenic capacity in the absence of a PCOS phenotype, suggesting a thecal defect induced during fetal life.

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Identification of MicroRNAs in Human Follicular Fluid: Characterization of MicroRNAs That Govern Steroidogenesis in Vitro and Are Associated With Polycystic Ovary Syndrome in Vivo

The Journal of Clinical Endocrinology & Metabolism ◽

10.1210/jc.2013-1715 ◽

2013 ◽

Vol 98 (7) ◽

pp. 3068-3079 ◽

Cited By ~ 152

Author(s):

Qing Sang ◽

Zhongyuan Yao ◽

Huan Wang ◽

Ruizhi Feng ◽

Haojue Wang ◽

...

Keyword(s):

Polycystic Ovary Syndrome ◽

Follicular Fluid ◽

Polycystic Ovary ◽

Ovary Syndrome ◽

Human Follicular Fluid ◽

Fluid Characterization

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A combination of polymorphisms in HSD11B1 associates with in vivo 11β-HSD1 activity and metabolic syndrome in women with and without polycystic ovary syndrome

Acta Endocrinologica ◽

10.1530/eje-11-0091 ◽

2011 ◽

Vol 165 (2) ◽

pp. 283-292 ◽

Cited By ~ 32

Author(s):

Alessandra Gambineri ◽

Federica Tomassoni ◽

Alessandra Munarini ◽

Roland H Stimson ◽

Roberto Mioni ◽

...

Keyword(s):

Metabolic Syndrome ◽

Adipose Tissue ◽

Polycystic Ovary Syndrome ◽

Polycystic Ovary ◽

Mrna Levels ◽

Nucleotide Polymorphisms ◽

Ovary Syndrome ◽

The Metabolic Syndrome ◽

Expression And Activity

ObjectiveRegeneration of cortisol by 11β-hydroxysteroid dehydrogenase type 1 (11β-HSD1) within liver and adipose tissue may be of pathophysiological importance in obesity and the metabolic syndrome. single nucleotide polymorphisms (SNPs) in HSD11B1, the gene encoding 11β-HSD1, have been associated with type 2 diabetes and hypertension in population-based cohort studies, and with hyperandrogenism in patients with the polycystic ovary syndrome (PCOS). However, the functional consequences of these SNPs for in vivo 11β-HSD1 expression and activity are unknown.MethodsWe explored associations of well-characterised hormonal and metabolic phenotypes with two common SNPs (rs846910 and rs12086634) in HSD11B1 in 600 women (300 with PCOS) and investigated 11β-HSD1 expression and activity in a nested study of 40 women from this cohort.ResultsHSD11B1 genotypes (as single SNPs and as the combination of the two minor allele SNPs) were not associated with PCOS. Women who were heterozygous for rs846910 A and homozygous for rs12086634 T (GA, TT genotype) had a higher risk of metabolic syndrome, regardless of the diagnosis of PCOS (odds ratio in the whole cohort=2.77 (95% confidence interval (CI) 1.16–6.67), P=0.023). In the nested cohort, women with the GA, TT genotype had higher HSD11B1 mRNA levels in adipose tissue, and higher rates of appearance of cortisol and d3-cortisol (16.1±0.7 nmol/min versus 12.1±1.1, P=0.044) during 9,11,12,12-2H4-cortisol (d4-cortisol) steady-state infusion.ConclusionsWe conclude that, in a population of Southern European Caucasian women with and without PCOS, alleles of HSD11B1 containing the two SNPs rs846910 A and rs12086634 T confer increased 11β-HSD1 expression and activity, which associates with the metabolic syndrome.

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