Huang Qun, Li Yuanbin, Chen Zhuang, Ou Huiping, Tan Yanjiao, Lin Hui
Department of Gynecology, The First Affiliated Hospital of Hunan Traditional Chinese Medical College, Zhuzhou, 412012, Hunan Province, People's Republic of China.
Department of Traditional Chinese Medicine, Hunan Traditional Chinese Medical College, No. 88 Zhihui Road, Shifeng District, Zhuzhou, 412012, Hunan Province, People's Republic of China.
J Ovarian Res. 2024 Feb 1;17(1):29. doi: 10.1186/s13048-024-01355-x.
Polycystic ovary syndrome (PCOS) is a frequent and complicated endocrine disease that remains a major reason for infertility. Bushenhuoluo Decotion (BSHLD) has been validated to exhibit curative effects on PCOS. This study was aimed to explore the potential mechanism underlying the therapeutic action of BSHLD.
PCOS rat model was induced by dehydroepiandrosterone (DHEA). Serum hormone and cytokines levels and ovarian pathological alterations were measured to assess ovarian function. Exosomes (Exos) were identified by Transmission electron microscopy and Nanoparticle Tracking Analysis. RT-qPCR, Western blotting, immunohistochemical staining, and immunofluorescence staining were performed to detect molecule expressions. Proliferation and pyroptosis of granulosa cells (GCs) were evaluated by CCK-8 and flow cytometry, respectively. The binding relationship between miR-30a-5p and suppressor of cytokine signaling 3 (SOCS3) was verified by dual luciferase reporter and RIP assays.
BSHLD treatment improved serum hormone abnormality, insulin sensitivity, and ovarian morphologic changes of PCOS rats. Moreover, BSHLD treatment restrained the excessive autophagy and pyroptosis in ovarian tissues of PCOS rats. Moreover, BSHLD reduced the expression of miR-30a-5p in serum, serum-derived Exos, and ovarian tissues, thus inhibiting autophagy and NLRP3-mediated pyroptosis in GCs. Mechanistically, SOCS3 was proved as a target of miR-30a-5p and could activate mTOR/P70S6K pathway to repress autophagy. The inhibitory effect of miR-30a-5p deficiency on autophagy and pyroptosis of GCs was attenuated by rapamycin.
Collectively, BSHLD suppressed autophagy and pyroptosis to improve POCS by regulating exosomal miR-30a-5p/SOCS3/mTOR signaling.
多囊卵巢综合征（PCOS）是一种常见且复杂的内分泌疾病，仍是不孕症的主要原因。补肾活络方（BSHLD）已被证实对PCOS有治疗作用。本研究旨在探讨BSHLD治疗作用的潜在机制。
用脱氢表雄酮（DHEA）诱导建立PCOS大鼠模型。检测血清激素和细胞因子水平以及卵巢病理改变以评估卵巢功能。通过透射电子显微镜和纳米颗粒跟踪分析鉴定外泌体（Exos）。进行RT-qPCR、蛋白质印迹、免疫组织化学染色和免疫荧光染色以检测分子表达。分别通过CCK-8和流式细胞术评估颗粒细胞（GCs）的增殖和焦亡。通过双荧光素酶报告基因和RIP试验验证miR-30a-5p与细胞因子信号传导抑制因子3（SOCS3）之间的结合关系。
BSHLD治疗改善了PCOS大鼠的血清激素异常、胰岛素敏感性和卵巢形态变化。此外，BSHLD治疗抑制了PCOS大鼠卵巢组织中过度的自噬和焦亡。此外，BSHLD降低了血清、血清来源的Exos和卵巢组织中miR-30a-5p的表达，从而抑制了GCs中的自噬和NLRP3介导的焦亡。机制上，证明SOCS3是miR-30a-5p的靶标，并且可以激活mTOR/P70S6K途径以抑制自噬。雷帕霉素减弱了miR-30a-5p缺乏对GCs自噬和焦亡的抑制作用。
总体而言，BSHLD通过调节外泌体miR-30a-5p/SOCS3/mTOR信号传导抑制自噬和焦亡，从而改善PCOS。
