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

Suqin Wang; Yingfang Wang; Qin Qin; Jianfang Li; Qiaoyun Chen; Ye Zhang; Xiuqing Li; Jianrong Liu

doi:10.1007/s12010-023-04825-y

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 Dec 28. doi: 10.1007/s12010-023-04825-y. Online ahead of print.

Authors

Suqin Wang^{1

2}, Yingfang Wang³, Qin Qin^{1

4}, Jianfang Li^{1

2}, Qiaoyun Chen^{1

4}, Ye Zhang^{1

4}, Xiuqing Li⁵, Jianrong Liu^{6

7}

Affiliations

¹ The Fifth Clinical Medical College of Shanxi Medical University, Taiyuan City, Shanxi, China.
² Department of Gynecology & Obstetrics, Fifth Hospital of Shanxi Medical University, Taiyuan City, 030012, Shanxi, China.
³ College of Basic Medicine and Forensic Medicine, Henan University of Science and Technology, Luoyang City, 471023, Henan, China.
⁴ Center for Reproductive Medicine, Fifth Hospital of Shanxi Medical University, No. 29, Shuangtasi Road, Yingze District, Taiyuan City, 030012, Shanxi, China.
⁵ Reproductive center, Coal Central Hospital of Shanxi Province, Taiyuan City, Shanxi, China.
⁶ The Fifth Clinical Medical College of Shanxi Medical University, Taiyuan City, Shanxi, China. liujianrong3@sxmu.edu.cn.
⁷ Center for Reproductive Medicine, Fifth Hospital of Shanxi Medical University, No. 29, Shuangtasi Road, Yingze District, Taiyuan City, 030012, Shanxi, China. liujianrong3@sxmu.edu.cn.

PMID: 38153651
DOI: 10.1007/s12010-023-04825-y

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.

Keywords: BBR; Circ_0097636; MiR-186-5p; PCOS; SIRT3.