二甲双胍通过PI3K/Akt/mTOR信号轴调控多囊卵巢综合征颗粒细胞自噬  

Metformin Regulates Autophagy of Granulosa Cells in Polycystic Ovary Syndrome through PI3K/Akt/mTOR Signal Axis

作　　者：刘艳萍 董伟 段伟静 高争 王晓红 LIU Yan-ping;DONG Wei;DUAN Wei-jing;GAO Zheng;WANG Xiao-hong(Department of Pharmacy,Ji'nan People's Hospital,Ji'nan,Shandong,271199,China;Department of Gynecology,Ji'nan People's Hospital,Jinan,Shandong,271199,China)

机构地区：[1]济南市人民医院药剂科,山东济南271199 [2]济南市人民医院妇科,山东济南271199

出　　处：《现代生物医学进展》2025年第7期1132-1141,共10页Progress in Modern Biomedicine

基　　金：山东省自然科学基金项目(ZR2021MH329);济南市科技局临床医学科技创新计划项目(202225067)。

摘　　要：目的:探讨二甲双胍调控多囊卵巢综合征(PCOS)颗粒细胞自噬的作用及潜在机制。方法:分离培养大鼠卵巢颗粒细胞并鉴定,随后分为对照组、模型组,低剂量组、高剂量组及联合组,除对照组外,其余各组采用丙酸睾酮+C2-神经酰胺建立PCOS卵巢颗粒细胞自噬模型,低剂量组、高剂量组及联合组分别添加0.5 mmol/L二甲双胍、1 mmol/L二甲双胍、1 mmol/L二甲双胍+10μmol/L的LY294002。噻唑蓝(MTT)法、流式细胞术、单丹磺尸酰胺(MDC)及Western blot法分别检测各组细胞增殖、凋亡、自噬及相关蛋白表达情况。结果:与对照组相比,模型组细胞24 h、48 h OD值减小,凋亡率和自噬率增升高,Beclin1表达及LC3Ⅱ/LC3Ⅰ均升高,p62、p-PI3K、p-Akt及p-mTOR表达均降低(P<0.05);与模型组比较,低剂量组和高剂量组24 h、48 h OD值增加,凋亡率和自噬率降低,Beclin1表达及LC3Ⅱ/LC3Ⅰ均降低,p62、p-PI3K、p-Akt及p-mTOR表达均升高(P <0.05),且高剂量组上述指标均优于低剂量组(P <0.05);相较于高剂量组,联合组细胞24 h、48 h OD值减小,凋亡率和自噬率增升高,Beclin1表达及LC3Ⅱ/LC3Ⅰ均升高,p62、p-PI3K、p-Akt及p-mTOR表达均降低(P <0.05)。结论:二甲双胍抑制PCOS卵巢颗粒细胞自噬和凋亡,促进细胞增殖,可能通过调控PI3K/AKT/mTOR信号通路发挥作用。Objective:To investigate the effect and potential mechanism of metformin on regulating autophagy in granulosa cells of polycystic ovary syndrome( PCOS).Methods:Rat ovarian granulosa cells were isolated and cultured,and identified.Subsequently,they were divided into control groups,model groups,low-dose group,high-dose group,and combined group.Except for the control group,the cells in other groups were to establish PCOS by using testosterone propionate + C2-ceramide.0.5 mmol/L metformin,1 mmol/L metformin,1 mmol/L metformin +10 μmol/L LY294002 were added to the low dose group,the high dose group and the combined group respectively.Cell proliferation,apoptosis,autophagy and related protein expression were detected by MTT assay,flow cytometry,monodansyl cadaverine( MDC) and Western blot.Results:Compared with that in the control group,the OD value of cells in the model group at 24 h and 48 h,and the expressions of p62,p-PI3K,p-Akt and p-mTOR decreased( P<0.05).While the apoptosis rate,autophagy rate increased,and the expressions of Beclin1and LC3 Ⅱ/LC3 Ⅰ increased.Compared with that in the model group,the 24 h and 48 h OD values of the low-dose group and the high-dose group increased,while the apoptosis rate and autophagy decreased.The expression of Beclin1 and LC3 Ⅱ/LC3 Ⅰ increased,and the expression of p62,p-PI3K,p-Akt and p-mTOR increased( P<0.05).The above indexes in the high-dose group were better than those in the low-dose group( P<0.05).Compared with the high-dose group,the OD value of cells in the combined group decreased at 24 h and 48 h,while the apoptosis rate,autophagy rateand the expressions of Beclin1and LC3 Ⅱ/LC3 Ⅰ increased,while the expressions of p62,p-PI3K,p-Akt and p-mTOR decreased( P<0.05).Conclusion:Metformin inhibits autophagy and apoptosis of ovarian granulosa cells in PCOS and promotes cell proliferation,which may play a role by regulating PI3K/AKT/mTOR signaling pathway.

关 键 词：多囊卵巢综合征 二甲双胍 颗粒细胞 自噬 凋亡 

分 类 号：R3[医药卫生—基础医学] R711.75