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Berberine Protects Against Dihydrotestosterone-Induced Human Ovarian Granulosa Cell Injury and Ferroptosis by Regulating the Circ_0097636/MiR-186-5p/SIRT3 Pathway

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Abstract

Polycystic ovarian syndrome (PCOS) is an endocrine syndrome in women of reproductive age. Berberine (BBR) is a Chinese herbal monomer that exhibits many pharmacological properties related to PCOS treatment. This study aims to analyze the effect of BBR on a cell model of PCOS and the underlying mechanism. Human ovarian granulosa (KGN) cells were treated with dihydrotestosterone (DHT) to mimic a PCOS cell model. The RNA expression of circ_0097636, miR-186-5p, and sirtuin3 (SIRT3) was determined by quantitative real-time polymerase chain reaction (qRT-PCR). Protein expression was detected by western blotting. Cell viability was analyzed by CCK-8 assay. Cell proliferation and apoptosis were investigated by 5-ethynyl-2′-deoxyuridine (EdU) assay and flow cytometry assay, respectively. The levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) were analyzed by enzyme-linked immunosorbent assays (ELISAs). Fe2+ concentration was assessed by an iron assay kit. Oxidative stress was assessed by detecting reactive oxygen species (ROS) level and malondialdehyde (MDA) level using commercial kits. The association of miR-186-5p with circ_0097636 and SIRT3 was identified by dual-luciferase reporter assay and RNA pull-down assay. Circ_0097636 expression was downregulated in the follicular fluid of PCOS patients and DHT-treated KGN cells when compared with control groups. BBR treatment partially relieved the DHT-induced inhibitory effect on cell proliferation and promoted effects on cell apoptosis, inflammation, ferroptosis, and oxidative stress in KGN cells. Additionally, circ_0097636 bound to miR-186-5p, and SIRT3 was identified as a target gene of miR-186-5p in KGN cells. BBR treatment ameliorated DHT-induced KGN cell injury by upregulating circ_0097636 and SIRT3 expression and downregulating miR-186-5p expression. Moreover, circ_0097636 overexpression protected KGN cells from DHT-induced injury by increasing SIRT3 expression. BBR ameliorated DHT-induced KGN cell injury and ferroptosis by regulating the circ_0097636/miR-186-5p/SIRT3 pathway.

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Funding

This work was supported by the Fund Program for the Scientific Activities of Selected Returned Overseas Professionals in Shanxi Province (No. 20230055) and the Fund Program for Research Project Supported by Shanxi Scholarship Council of China (No. 2023-183).

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Authors and Affiliations

  1. The Fifth Clinical Medical College of Shanxi Medical University, Taiyuan City, Shanxi, China

    Suqin Wang, Qin Qin, Jianfang Li, Qiaoyun Chen, Ye Zhang & Jianrong Liu

  2. Department of Gynecology & Obstetrics, Fifth Hospital of Shanxi Medical University, Taiyuan City, 030012, Shanxi, China

    Suqin Wang & Jianfang Li

  3. College of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang City, 471023, Henan, China

    Yingfang Wang

  4. Center for Reproductive Medicine, Fifth Hospital of Shanxi Medical University, No. 29, Shuangtasi Road, Yingze District, Taiyuan City, 030012, Shanxi, China

    Qin Qin, Qiaoyun Chen, Ye Zhang & Jianrong Liu

  5. Reproductive center, Coal Central Hospital of Shanxi Province, Taiyuan City, Shanxi, China

    Xiuqing Li

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Contributions

SW designed and performed the research; YW, QQ, JfL, QC, YZ, XL, and JrL analyzed the data; SW wrote the manuscript. All authors read and approved the final manuscript.

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Correspondence to Jianrong Liu.

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Wang, S., Wang, Y., Qin, Q. et al. Berberine Protects Against Dihydrotestosterone-Induced Human Ovarian Granulosa Cell Injury and Ferroptosis by Regulating the Circ_0097636/MiR-186-5p/SIRT3 Pathway. Appl Biochem Biotechnol (2023). https://doi.org/10.1007/s12010-023-04825-y

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