新加苁蓉菟丝汤通过FtMt/PINK1/Parkin调节人卵巢颗粒细胞线粒体铁超载抗铁死亡的分子机制OA
The molecular mechanism of Xinjia Congrong Tusi Decoction(新加苁蓉菟丝汤)in regulating mitochondrial iron overload to inhibit ferroptosis in human ovarian granulosa cells via the FtMt/PINK1/Parkin pathway
目的 探究新加苁蓉菟丝汤(XCTD)通过FtMt/PINK1/Parkin调节人卵巢颗粒细胞线粒体铁超载,修复雷公藤甲素(triptolide,TP)致卵巢颗粒细胞功能减退及损伤、抗铁死亡,促进卵泡发育的分子机制.方法 将体外培养的人卵巢颗粒细胞(granulosa cells,GCs)KGN细胞分为7组:A组:正常对照组(Control);B组:TP模型组(TP);C组:TP+10%含药血清新加苁蓉菟丝汤剂量组(TP+XCTD);D组:TP+西药组(TP+EV);E组:新加苁蓉菟丝汤+铁死亡激动剂组(TP+XCTD+erastin).F组:新加苁蓉菟丝汤+铁死亡激动剂+OE-NC组(TP+XCTD+erastin+NC);G组:新加苁蓉菟丝汤+铁死亡激动剂+OE-FtMt组(TP+XCTD+erastin+FtMt).以上各组采用流式细胞仪检测人卵巢颗粒细胞中脂质过氧化情况;透射电镜观察各组人卵巢颗粒细胞中铁死亡相关细胞器形态变化;荧光染核实验观察各组颗粒细胞及线粒体变化;生化检测各组颗粒细胞线粒体总铁含量和GSH表达;Western blot法检测各组卵巢颗粒细胞中FtMt、PINK1、Parkin、SLC7A11、GPX4的蛋白表达量.结果 与B组相比,C组、D组和G组颗粒细胞脂质过氧化水平显著降低;与B组相比,C组、D组和G组电镜下颗粒细胞形态和结构恢复较好;与B组相比,C组、D组和G组颗粒细胞荧光强度均显著升高;通过生化检测发现,C组、D组和G组较B组颗粒细胞中线粒体铁含量明显降低而GSH水平显著升高;经过Western blot检测发现,与B组相比,C组、D组和G组颗粒细胞中FtMt、SLC7A11、GPX4的蛋白表达量升高而PINK1、Parkin蛋白表达量降低.结论 新加苁蓉菟丝汤可能通过调节FtMt/PINK1/Parkin通路,改善人卵巢颗粒细胞线粒体铁超载,从而逆转颗粒细胞铁死亡.
Objective To explore the molecular mechanism of Xinjia Congrong Tusi Decoction(新加苁蓉菟丝汤,XCTD)in regulat-ing mitochondrial iron overload via the FtMt/PINK1/Parkin pathway,repairing the triptolide(TP)-induced dysfunction and damage of human ovarian granulosa cells(GCs),inhibiting ferroptosis,and promoting follicular development.Methods Cultured human ovarian granulosa cells(KGN cells)were divided into 7 groups:Group A:normal control(Control);Group B:TP model(TP);Group C:TP+10%drug serum XCTD(TP+XCTD);Group D:TP+Western medicine(TP+EV);Group E:XCTD+ferroptosis agonist(TP+XCTD+erastin);Group F:XCTD+ferroptosis agonist+NC(TP+XCTD+erastin+NC);Group G:XCTD+ferroptosis agonist+FtMt(TP+XCTD+erastin+FtMt).Lipid peroxidation in GCs was detected by flow cytometry;morphological changes of ferroptosis-related organelles in GCs were observed by transmission electron microscopy(TEM);changes in GCs and mitochondria were observed by nuclear fluorescence staining;total mitochondrial iron content and GSH expression in GCs were measured biochemically;and protein expression levels of FtMt,PINK1,Parkin,SLC7A11,and GPX4 in GCs were determined by Western blot(WB).Results Compared with Group B,lipid peroxidation levels in GCs were significantly lower in Groups C,D,and G.Compared with Group B,the morphol-ogy and structure of GCs recovered better under electron microscopy in Groups C,D,and G.Compared with Group B,nuclear fluores-cence intensity in GCs was significantly higher in Groups C,D,and G.Biochemical assays showed that mitochondrial iron content in GCs was significantly lower,while GSH levels were significantly higher in Groups C,D,and G compared to Group B.WB analysis revealed that,compared with Group B,the protein expression levels of FtMt,SLC7A11,and GPX4 in GCs were increased,while the levels of PINK1 and Parkin were decreased in Groups C,D,and G.Conclusion XCTD may inhibit ferroptosis in human ovarian granu-losa cells by regulating the FtMt/PINK1/Parkin pathway,thereby ameliorating mitochondrial iron overload.
严谨;吴克明;张惜燕
陕西中医药大学基础医学院,陕西 咸阳 712046成都中医药大学附属医院,四川 成都 610000陕西省中医体质与疾病防治重点实验室,陕西 咸阳 712046
医药卫生
新加苁蓉菟丝汤卵巢功能减退FtMt/PINK1/Parkin通路铁死亡颗粒细胞
Xinjia Congrong Tusi Decoction(新加苁蓉菟丝汤)Ovarian hypofunctionFtMt/PINK1/Parkin PathwayFerroptosisGranulosa cells
《时珍国医国药》 2026 (17)
3201-3207,7
国家自然科学基金(82405465)陕西省科技厅科研计划一般项目(2024JC-YBQN-0892)陕西省教育厅重点科研计划项目(21JS017)
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