miR-215-5p调控YY1通过TGF-β1/Smad信号通路对PCOS大鼠卵巢颗粒细胞凋亡的影响  

Influence of miR-215-5p on apoptosis of ovarian granulosa cells in rats with polycystic ovary syndrome by regulating YY1 through TGF-β1/Smad signaling pathway

作　　者：陈莹莹 齐小雪[1] 苟元钦 CHEN Yingying;QI Xiaoxue;GOU Yuanqin(Department of Obstetrics and Gynecology,Chengdu First People's Hospital,Chengdu 610041,China)

机构地区：[1]成都市第一人民医院妇产科,四川成都610041

出　　处：《西部医学》2023年第5期662-669,共8页Medical Journal of West China

基　　金：四川省医学会专项科研项目(2019HR65)。

摘　　要：目的探究MicroRNA-215-5p(miR-215-5p)在多囊卵巢综合征(PCOS)大鼠中的表达及其对卵巢颗粒细胞(GC)凋亡的影响,并分析潜在机制。方法采用脱氢表雄酮(DHEA)建立PCOS大鼠模型,分离并培养原代PCOS大鼠卵巢GC,分为对照组(control组)、sh-NC组、sh-YY1组、sh-YY1+LY2109761组、miR-NC组、miR-215-5p mimic组、miR-215-5p mimic+pc-NC组、miR-215-5p mimic+pc-YY1组。TUNEL法检测PCOS大鼠卵巢组织GC凋亡;实时荧光定量PCR(RT-qPCR)检测卵巢组织/GC中miR-215-5p、阴阳-1(YY1)mRNA表达;CCK-8法和平板克隆形成实验检测细胞增殖能力;流式细胞仪检测细胞凋亡率;Western blot检测PCOS大鼠卵巢组织/GC中YY1、转化生长因子-β1(TGF-β1)、Smad4蛋白表达。结果miR-215-5p在PCOS大鼠卵巢组织中低表达,YY1 mRNA和蛋白水平在PCOS大鼠卵巢组织中显著升高(P<0.05);敲低YY1或上调miR-215-5p可显著增强PCOS大鼠卵巢GC增殖能力,抑制GC凋亡,升高TGF-β1、Smad4蛋白水平(均P<0.05);TGF-β/Smad4信号通路抑制剂LY2109761可显著减弱敲低YY1对卵巢GC凋亡的抑制作用(P<0.05);在过表达miR-215-5p的基础上,上调YY1可抑制TGF-β1/Smad信号通路的激活,显著减弱miR-215-5p mimic对PCOS大鼠卵巢GC凋亡的抑制作用(P<0.05)。结论miR-215-5p在PCOS中呈低表达,过表达miR-215-5p可能通过下调YY1,激活TGF-β1/Smad信号通路,促进PCOS大鼠卵巢GC增殖并抑制其凋亡。Objective To investigate the expression of MicroRNA-215-5p(miR-215-5p)in polycystic ovary syndrome(PCOS)rats and its influence on ovarian granulosa cell(GC)apoptosis,and analyze the underlying mechanism.Methods Dehydroepiandrosterone(DHEA)was used to establish a PCOS rat model,and primary PCOS rat ovarian GCs were isolated and cultured.They were grouped into control group,sh-NC group,sh-YY1 group,sh-YY1+LY2109761 group,miR-NC group,miR-215-5p mimic group,miR-215-5p mimic+pc-NC group,and miR-215-5p mimic+pc-YY1 group.The GC apoptosis in PCOS rat ovarian tissue was detected by TUNEL method;the mRNA expression of miR-215-5p and Yin-Yang-1(YY1)in ovarian tissue/GC was measured by real-time quantitative PCR(RT-qPCR);cell proliferation ability was detected by CCK-8 method and plate colony formation assay;cell apoptosis rate was measured by flow cytometry;the protein expression of YY1,transforming growth factor-β1(TGF-β1)and Smad4 in ovarian tissue/GC of PCOS rats was detected by Western blot.Results The expression of miR-215-5p was low in the ovarian tissue of PCOS rats,and the mRNA and protein levels of YY1 were significantly increased in the ovarian tissue of PCOS rats(P<0.05).Knockdown of YY1 or up-regulation of miR-215-5p was able to significantly enhance the proliferation of ovarian GCs in PCOS rats,inhibit GC apoptosis,and increase the levels of TGF-β1 and Smad4 proteins(all P<0.05).TGF-β/Smad4 signaling pathway inhibitor LY2109761 was able to significantly attenuate the inhibitory effect of knockdown of YY1 on ovarian GC apoptosis(P<0.05);on the basis of overexpression of miR-215-5p,up-regulation of YY1 was able to inhibit the activation of TGF-β1/Smad signaling pathway,and significantly attenuate the inhibitory effect of miR-215-5p mimic on ovarian GC apoptosis in PCOS rats(P<0.05).Conclusion MiR-215-5p is lowly expressed in PCOS.Overexpression of miR-215-5p may promote the proliferation of ovarian GC and inhibit apoptosis in PCOS rats by downregulating YY1 and activating the TGF-β1/Smad signaling pathway

关 键 词：多囊卵巢综合征 MicroRNA-215-5p 卵巢颗粒细胞 凋亡 阴阳-1 转化生长因子-β1/Smad信号通路 

分 类 号：R711.75[医药卫生—妇产科学]