SPARC介导卵巢颗粒细胞凋亡经由Wnt/β-catenin信号通路  

SPARC-mediated apoptosis of ovarian granulosa cells via the Wnt/β-catenin signaling pathway

作　　者：魏艳会 党洁 WEI Yanhui;DANG Jie(Department of Obstetrics and Gynecology,Shaanxi Nuclear Industry 215 Hospital,Xianyang,Shaanxi 712000,China)

机构地区：[1]陕西省核工业二一五医院产科/妇科,陕西咸阳712000

出　　处：《中国优生与遗传杂志》2024年第12期2501-2507,共7页Chinese Journal of Birth Health & Heredity

摘　　要：目的本研究旨在探讨富含半胱氨酸的酸性分泌蛋白(SPARC)经由Wnt/β-catenin通路介导卵巢颗粒细胞凋亡,初探其对多囊卵巢综合征(PCOS)的研究意义。方法通过转染技术过表达和敲低人卵巢颗粒细胞KGN中的SPARC蛋白,并采用RT-qPCR法验证转染效率;采用CCK-8法检测细胞增殖能力;采用TUNEL法检测细胞凋亡率;采用蛋白质印迹法检测Wnt/β-catenin通路相关蛋白表达水平;采用酶联免疫吸附试验(ELISA)检测类固醇激素雌二醇(E_(2))和孕酮(PG)的分泌水平;并使用Wnt/β-catenin通路抑制剂WIKI4验证SPARC与Wnt/β-catenin通路的关系。结果SPARC过表达可显著提高颗粒细胞Wnt、β-catenin和p-Foxo3a的蛋白表达水平,降低Foxo3a表达,促进细胞增殖并减少凋亡率。SPARC表达下调则表现出相反的效果。WIKI4处理证实了SPARC通过Wnt/β-catenin通路调控颗粒细胞的增殖和凋亡。此外,SPARC过表达提高了E_(2)和PG的分泌水平,而其低表达则降低了这些激素的分泌。结论本研究揭示了SPARC通过Wnt/β-catenin信号通路调控卵巢颗粒细胞凋亡的机制,为PCOS的病理机制研究提供了新视角,并指出SPARC可能作为治疗PCOS的潜在靶点。Objective This study aimed to investigate the secreted protein acidic and rich in cysteine(SPARC)-mediated ovarian granulosa cell apoptosis via the Wnt/β-catenin pathway and to first explore its significance in polycystic ovary syndrome(PCOS).Methods The SPARC protein was overexpressed and knocked down in human ovarian granulosa cells KGN by transfection,and the transfection efficiency was verified by RT-qPCR.Cell proliferation ability was detected by CCK-8.Apoptosis rate was detected by TUNEL assay.Wnt/β-catenin pathway related proteins were detected by Western blotting.And steroids and estradiol(E_(2))were detected by ELISA.The secretion levels of estradiol(E_(2))and progesterone(PG)were detected by ELISA,and the relationship between SPARC and the Wnt/β-catenin pathway was verified by using the Wnt/β-catenin pathway inhibitor WIKI4.Results SPARC overexpression significantly increased the protein expression levels of Wnt,β-catenin,and p-Foxo3a and decreased Foxo3a expression in granulosa cells,promoting cell proliferation and decreasing apoptosis rates.Down-regulation of SPARC expression showed the opposite WIKI4 treatment confirmed that SPARC regulates granulosa cell proliferation and apoptosis through the Wnt/β-catenin pathway.Proliferation and apoptosis.WIKI4 treatment confirmed that SPARC regulates granulosa cell proliferation and apoptosis through the Wnt/β-catenin pathway.Proliferation and apoptosis.In addition,SPARC overexpression increased the secretion levels of E_(2) and PG,whereas its lower expression decreased the Wnt/β-cateninecretion of these hormones.Conclusion This study revealed the mechanism of SPARC regulating ovarian granulosa cell apoptosis through the Wnt/β-catenin signaling pathway,which provides a new perspective for the study of the pathological mechanism of PCOS and indicates that SPARC may serve as a potential target for the treatment of PCOS.

关 键 词：富含半胱氨酸的酸性分泌蛋白 卵巢颗粒细胞 WNT/Β-CATENIN通路 凋亡 多囊卵巢综合征 

分 类 号：R73[医药卫生—肿瘤]