桂枝茯苓丸含药血清对多囊卵巢综合征小鼠卵巢颗粒细胞自噬的影响  

Effects of Guizhi Fuling Pills medicated serum on autophagy of ovarian granulosa cells in mice with polycystic ovary syndrome

作　　者：陈海燕[1] 朱鸿秋[2] 朱影 胡小丹 吴沛娟 吴晗雪 CHEN Hai-yan;ZHU Hong-qiu;ZHU Ying;HU Xiao-dan;WU Pei-juan;WU Han-xue(Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China;School of Medical and Life Sciences,Chengdu University of Traditional Chinese Medicine,Chengdu 610072,China)

机构地区：[1]成都中医药大学,四川成都610072 [2]成都中医药大学医学与生命科学院,四川成都610072

出　　处：《中成药》2022年第10期3137-3144,共8页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金项目(81873335);四川省科学技术厅项目基金(2020YJ0376);成都中医药大学医学与生命科学院医院专项基金项目(2020yky05)。

摘　　要：目的 探讨桂枝茯苓丸含药血清通过调控基因靶点H19/miR-29b-3p对多囊卵巢综合征(PCOS)小鼠卵巢颗粒细胞自噬的影响。方法 分离及培养PCOS小鼠卵巢颗粒细胞,转染oe-H19或miR-29b-3p-inhibitor,使用桂枝茯苓丸含药血清进行干预,采用流式细胞仪和Western blot法分别检测卵巢颗粒细胞自噬率及LC3Ⅱ/Ⅰ蛋白表达,RT-qPCR法检测卵巢颗粒细胞H19、miR-29b-3p mRNA表达。结果 与对照组比较,模型组小鼠卵巢颗粒细胞自噬率、LC3Ⅱ/Ⅰ蛋白和H19 mRNA表达升高(P<0.01),miR-29b-3p mRNA表达降低(P<0.01);与模型组比较,桂枝茯苓丸含药血清组卵巢颗粒细胞自噬率及LC3Ⅱ/Ⅰ蛋白表达均降低(P<0.01),H19、miR-29b-3p mRNA表达均发生逆转(P<0.01);过表达H19或抑制miR-29b-3p表达可减弱桂枝茯苓丸含药血清对卵巢颗粒细胞过度自噬的抑制作用(P<0.01)。结论 桂枝茯苓丸含药血清可以通过H19/miR-29b-3p途径抑制PCOS小鼠卵巢颗粒细胞的自噬水平。AIM To investigate the effects of Guizhi Fuling Pills medicated serum on autophagy of ovarian granulosa cells in polycystic ovary syndrome(PCOS) mouse models via regulation of gene target H19/miR-29b-3p.METHODS The isolated and cultured ovarian granulosa cells of PCOS mouse models were intervened with Guizhi Fuling Pills medicated serum following transfection with oe-H19 or miR-29b-3p-inhibitor. Subsequently, the ovarian granulosa cells had their autophagy rate and LC3Ⅱ/Ⅰ protein expression detected by flow cytometry and Western blot, respectively;and their mRNA expressions of H19 and miR-29b-3p determined by RT-qPCR.RESULTS Compared with those of the control group, ovarian granulosa cells of the model group displayed increased autophagy rate and LC3Ⅱ/Ⅰ protein and H19 mRNA expressions(P<0.01), and decreased miR-29b-3p mRNA expression(P<0.01). Compared with the model group, the group dosed with Guizhi Fuling Pills medicated serum demonstrated decreased autophagy rate and LC3Ⅱ/Ⅰ protein expression(P<0.01), and reversed mRNA expressions of H19 and miR-29b-3p(P<0.01). Overexpression of H19 or inhibition of miR-29b-3p expression attenuated the inhibitory effect due to Guizhi Fuling Pills medicated serum on excessive autophagy in ovarian granulosa cells(P<0.01).CONCLUSION Guizhi Fuling Pills medicated serum can inhibit the autophagy level of ovarian granulosa cells in PCOS mice via H19/miR-29b-3p pathway.

关 键 词：桂枝茯苓丸 多囊卵巢综合征(PCOS) 卵巢颗粒细胞 自噬 H19/miR-29b-3p 

分 类 号：R285.5[医药卫生—中药学]