Addressing the role of 11β-hydroxysteroid dehydrogenase type 1 in the development of polycystic ovary syndrome and the putative therapeutic effects of its selective inhibition in a preclinical model

Li, Xinyu; Hu, Shuanggang; Zhu, Qinling; Yao, Guangxin; Yao, Jun; Li, Jiaxing; Wei, Yuan; Ding, Ying; Jia, Qi; Xu, Rui; Zhao, Hanting; Zhu, Zhenyi; Du, Yanzhi; Sun, Kang; Sun, Yun

doi:10.1016/j.metabol.2021.154749

Cited by 13 publications

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References 43 publications

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“…Animal experiments were approved by the Animal Care Committee of the Ren Ji Hospital, Shanghai Jiao Tong University School of Medicine. To establish the PCOS rat model, 3-week-old female SD rats were injected daily (s.c.) with DHEA (6 mg/100 g body weight (BW)) (Langchem, Shanghai, China) dissolved in 0.2 ml PBS for 20 consecutive days, after 1 week of adaptation, according to Li et al (2021b) . Meanwhile, the rats in the control group were injected with an equivalent volume of PBS.…”

Section: Methodsmentioning

confidence: 99%

Ovarian ferroptosis induced by androgen is involved in pathogenesis of PCOS

Li,

Lin,

Cheng

et al. 2024

Human Reproduction Open

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STUDY QUESTION Does ovarian ferroptosis play an active role in the development of PCOS? SUMMARY ANSWER Increased ovarian ferroptosis was present in PCOS ovaries and the inhibition of ferroptosis with ferrostatin-1 (Fer-1) ameliorated polycystic ovary morphology and anovulation. WHAT IS KNOWN ALREADY Programmed cell death plays a fundamental role in ovarian follicle development. However, the types and mechanisms of cell death involved in the ovary are yet to be elucidated. Ferroptosis is a recently discovered iron-dependent programmed cell death. Impaired iron metabolism and cell death have been observed in women with PCOS, the main cause of anovulatory infertility. Additionally, previous studies reported that an abnormal expression of noncoding RNA may promote ferroptosis in immortalized ovarian granulosa cell lines. However, little is known about whether ovarian ferroptosis is increased in PCOS, and there is insufficient direct evidence for a role of ferroptosis in PCOS, and the underlying mechanism. Moreover, the effect of the inhibition of ferroptosis with Fer-1 in PCOS remains unclear. STUDY DESIGN, SIZE, DURATION Ferroptosis was evaluated in human granulosa cells (hGCs) from non-PCOS （n = 6–16）and PCOS (n = 7–18) patients. The experimental study was completed in vitro using primary hGCs from women undergoing IVF. Improvements in PCOS indicators following ferroptosis inhibition with Fer-1 were investigated in a dehydroepiandrosterone (DHEA)-induced PCOS rat models (n = 8 per group). PARTICIPANTS/MATERIALS, SETTING, METHODS Ovarian ferroptosis was evaluated in the following ways: by detecting iron concentrations via ELISA and fluorescent probes; measuring malondialdehyde (MDA) concentrations via ELISA; assessing ferroptosis-related protein abundance with western blotting; observing mitochondrial morphology with transmission electron microscopy; and determining cell viability. Primary hGCs were collected from women undergoing IVF. They were treated with dihydrotestosterone (DHT) for 24 h. The effect of DHT on ferroptosis was examined in the presence or absence of small interfering RNA-mediated knockdown of the putative receptor coregulator for signalling molecules. The role of ovarian ferroptosis in PCOS progression was explored in vivo in rats. The DHEA-induced PCOS rat model was treated with the ferroptosis inhibitor, Fer-1, and the oocytes and metaphase II (MII) oocytes were counted after ovarian stimulation. Additionally, rats were treated with the ferroptosis inducer, RSL3, to further explore the effect of ferroptosis. The concentrations of testosterone, FSH, and LH were assessed. MAIN RESULTS AND THE ROLE OF CHANCE Increased ferroptosis was detected in the ovaries of patients with PCOS and in rats with DHEA-induced PCOS. Increased concentrations of Fe2+ (p < 0.05) and MDA (p < 0.05), and upregulated nuclear receptor coactivator 4 (NCOA4) protein levels, and downregulated ferritin heavy chain 1 (FTH1) and glutathione peroxidase 4 (GPX4) proteins were observed in the hGCs in patients with PCOS and ovaries of PCOS rats (p < 0.05 versus control). DHT was shown to induce ferroptosis via activation of NOCA4-dependent ferroptinophagy. The inhibition of ferroptosis with Fer-1 in rats ameliorated a cluster of PCOS traits including impaired glucose tolerance, irregular estrous cycles, reproductive hormone dysfunction, hyperandrogenism, polycystic ovaries, anovulation, and oocyte quality (p < 0.05). Treating rats with RSL3 resulted in polycystic ovaries and hyperandrogenism (p < 0.05). LARGE-SCALE DATA N/A. LIMITATIONS, REASONS FOR CAUTION Although ovarian-targeted ferroptosis inhibition may be a more targeted treatment for PCOS, the underlying mechanisms in the cycle between ferroptosis and hyperandrogenism requires further exploration. Additionally, since PCOS shows high heterogeneity, it is important to investigate whether ferroptosis increases are present in all patients with PCOS. WIDER IMPLICATIONS OF THE FINDINGS Androgen-induced ovarian ferroptosis appears to play a role in the pathogenesis of PCOS, which potentially makes it a promising treatment target in PCOS. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by the National Key R＆D Program of China (2023YFC2705500, 2023YFC2705505, 2019YFA0802604), National Natural Science Foundation of China (No. 82130046, 82320108009, 82101708, 82101747 and 82001517), Shanghai leading talent program, Innovative research team of high-level local universities in Shanghai (No. SHSMU-ZLCX20210201, No. SSMU-ZLCX20180401), Shanghai Jiaotong University School of Medicine Affiliated Renji Hospital Clinical Research Innovation Cultivation Fund Program (RJPY-DZX-003) and Shanghai Municipal Education Commission-Gaofeng Clinical Medicine Grant Support (No. 20161413), Shanghai's Top Priority Research Center Construction Project (2023ZZ02002) and Three-Year Action Plan for Strengthening the Construction of the Public Health System in Shanghai (GWVI-11.1-36). The authors report no competing interests.

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Section: Methodsmentioning

confidence: 99%

Ovarian ferroptosis induced by androgen is involved in pathogenesis of PCOS

Li,

Lin,

Cheng

et al. 2024

Human Reproduction Open

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“…A total of 54 female SD rats, aged three weeks, were chosen and assigned randomly to three groups: the DHEA group (n = 18), the oil control group (n = 18), and the blank control group (n = 18). The PCOS rat model was produced in Sprague-Dawley rats using the dehydroepiandrosterone (DHEA) technique [17].…”

Section: Pcos Model Constructionmentioning

confidence: 99%

AMH regulates ovarian granulosa cell growth in PCOS rats through SMAD4

Dong,

Liu,

Zhang

et al. 2024

Preprint

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Background Polycystic Povary syndrome(PCOS) is a diverse condition with an unknown cause. Anti-Mullerian hormone(AMH) is a hormone that belongs to the transforming growth factor-β(TGF-β) class. Mothers against decapentaplegic homolog 4(SMAD4) is a crucial transcription factor widely expressed in granulosa cells in the TGF-β signaling pathway. Previous studies have revealed that AMH may be an important factor in follicular developmental disorders in PCOS patients , as a biomarker of PCOS. Objective This study examines the involvement of AMH in the formation of ovulatory abnormalities in PCOS rats and explores its potential causes. Methods A PCOS rat model was created by inducing DHEA, and granulosa cells from the ovaries were extracted and identified. The expression of AMH and SMAD4 in PCOS rats was assessed by ELISA, immu-nohistochemistry, and Western blot; and the effects of different concentrations of AMH recombinant proteins on the expression of SMAD4 and the development of granulosa cells were examined. The effect of knocking down SMAD4 expression with siRNA on granulosa cell development was also examined. Results The expression of AMH and SMAD4 in the ovarian tissues and granulosa cells in the PCOS group was higher (*P<0.05). The expression of PCNA in the ovarian granulosa cells of the PCOS group was lower (**P<0.01), and the expression of BAX was higher (*P<0.05). Western blot analysis indicated that 100 ng/ml rAMH increased the expression of SMAD4 and caspase-3 in granulosa cells (*P<0.05), and decreased CyclinA and BCL-2 expression (**P<0.01). CCK-8 and flow cytometry results showed that 100 ng/ml AMH reduced proliferation and increased apoptosis in granulosa cells (***P<0.001,*P<0.05). siRNA knockdown of the SMAD4 gene led to increased PCNA expression (**P<0.01) and decreased BAX expression (*P<0.05) in granulosa cells of PCOS rats. Conclusions AMH plays a role in controlling the growth and programmed cell death of ovarian granulosa cells in rats with polycystic ovary syndrome through SMAD4.

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“…Rats in the model group were induced with DOR by intraperitoneal injection of 90 mg/kg cyclophosphamide (CTX; Beyotime) once [22]. Fourteen days after CTX administration, the DOR rats were gavaged with dimethyl sulfoxide (DMSO; Beyotime) or different doses of RD dissolved in DMSO (low: 19 mg/kg; medium: 38 mg/kg; high: 76 mg/kg) once a day for consecutive 2 weeks [18], during which lentiviral vectors of sh-FOXO1, sh-NC, LV-NC, or LV-KLOTHO (20 µl, virus titer: 10 9 TU/ml) were additionally injected into both ovaries of the DOR rats according to experimental design [23,24]. Fourteen days after injection of normal saline, sham-operated rats were intragastrically administrated with DMSO.…”

Section: A Rat Model Of Dormentioning

confidence: 99%

Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

Yan

Wang

Chen

et al. 2023

Korean J Physiol Pharmacol

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This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E 2 ) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E 2 expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.

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Addressing the role of 11β-hydroxysteroid dehydrogenase type 1 in the development of polycystic ovary syndrome and the putative therapeutic effects of its selective inhibition in a preclinical model

Cited by 13 publications

References 43 publications

Ovarian ferroptosis induced by androgen is involved in pathogenesis of PCOS

Ovarian ferroptosis induced by androgen is involved in pathogenesis of PCOS

AMH regulates ovarian granulosa cell growth in PCOS rats through SMAD4

Rehmannioside D mitigates disease progression in rats with experimental-induced diminished ovarian reserve via Forkhead Box O1/KLOTHO axis

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