沉默信息调节因子1/叉头框蛋白O1信号通路调控铁死亡对多囊卵巢综合征大鼠卵巢颗粒细胞的影响实验研究  

Effect of silent information regulator 1/forkhead box protein O1 signaling pathway on ovarian granulosa cells in rats with polycystic ovary syndrome by regulating ferroptosis

作　　者：杨加宁 张立然 YANG Jianing;ZHANG Liran(Heilongjiang Academy of Traditional Chinese Medicine,Harbin 150036,China)

机构地区：[1]黑龙江省中医药科学院,黑龙江哈尔滨150036 [2]黑龙江省中医药科学院南岗分院妇科,黑龙江哈尔滨150080

出　　处：《陕西医学杂志》2024年第8期1035-1040,共6页Shaanxi Medical Journal

基　　金：黑龙江省中医药科研项目(ZHY2023-082)。

摘　　要：目的:探讨沉默信息调节因子1(SIRT1)/叉头框蛋白O1(FOXO1)信号通路调控铁死亡对多囊卵巢综合征(PCOS)大鼠卵巢颗粒细胞的影响。方法:将30只雌性SD大鼠分为对照组(皮下注射豆油)和PCOS组(皮下注射脱氢表雄酮)。将对照组和PCOS组大鼠分离得到的卵巢颗粒细胞分为对照组、PCOS组,取一半PCOS组卵巢颗粒细胞为过表达SIRT1组(20μmol/L SIRT1激活剂SRT 2183干预)。RT-qPCR及Western blot检测细胞中SIRT1、FOXO1、溶质载体家族7成员11(SLC7A11)、谷胱甘肽过氧化物酶4(GPX4)、Bcl-2关联X蛋白(Bax)、半胱氨酸蛋白酶-3(Caspase-3)、B淋巴细胞瘤-2(Bcl-2)mRNA及蛋白表达。Hoechst染色检测细胞活性氧(ROS)及线粒体活性氧(mtROS)水平。ELISA检测细胞中谷胱甘肽(GSH)、丙二醛(MDA)、Fe^(2+)、卵泡刺激素(FSH)、睾酮(T)、黄体生成素(LH)水平。流式细胞术检测细胞凋亡情况。结果:与对照组比较,PCOS组卵巢颗粒细胞中SIRT1、FOXO1、SLC7A11、GPX4、Bcl-2 mRNA及蛋白表达和GSH、FSH水平降低,Bax、Caspase-3 mRNA及蛋白表达和MDA、ROS、mtROS、Fe^(2+)、T、LH水平升高,卵巢颗粒细胞凋亡率增加(均P<0.05)。与PCOS组比较,过表达SIRT1组卵巢颗粒细胞中SIRT1、FOXO1、SLC7A11、GPX4、Bcl-2 mRNA及蛋白表达和GSH、FSH水平升高,Bax、Caspase-3 mRNA及蛋白表达和MDA、ROS、mtROS、Fe^(2+)、T、LH水平降低,卵巢颗粒细胞凋亡率减少(均P<0.05)。结论:SIRT1/FOXO1通路在PCOS卵巢颗粒细胞中下调,激活该通路可能通过下调铁死亡抑制PCOS卵巢颗粒细胞凋亡。Objective:To investigate the effect of silent information regulator 1(SIRT1)/forkhead box protein O1(FOXO1)signaling pathway on ovarian granulosa cells in rats with polycystic ovary syndrome(PCOS)by regulating ferroptosis.Methods:Thirty female SD rats were divided into control group(subcutaneous injection of soybean oil)and PCOS group(subcutaneous injection of DHEA).The ovarian granulosa cells isolated from the control group and the PCOS group were divided into the control group and the PCOS group.Half of the ovarian granulosa cells in the PCOS group were selected as the over-expressing SIRT1 group(20μmol/L SIRT1 activator SRT 2183 intervention).The mRNA and protein expression of SIRT1,FOXO1,solute carrier family 7 member 11(SLC7A11),glutathione peroxidase 4(GPX4),Bcl-2-associated X protein(Bax),Caspase-3,B lymphoblastoma-2(Bcl-2)were detected by RT-qPCR and Western blot.The levels of reactive oxygen species(ROS)and mitochondrial ROS(mtROS)were detected by Hoechst staining.The levels of glutathione(GSH),malondialdehyde(MDA),Fe^(2+),follicle-stimulating hormone(FSH),testosterone(T)and luteinizing hormone(LH)were detected by ELISA.The apoptosis was detected by flow cytometry.Results:Compared with the control group,the mRNA and protein expressions of SIRT1,FOXO1,SLC7A11,GPX4,Bcl-2 and the levels of GSH and FSH in ovarian granulosa cells of PCOS group were decreased,while the mRNA and protein expressions of Bax and Caspase-3 and the levels of MDA,ROS,Fe^(2+),T and LH were increased(all P<0.05).The apoptosis rate of ovarian granulosa cells was increased(P<0.05).Compared with PCOS group,mRNA and protein expressions of SIRT1,FOXO1,SLC7A11,GPX4,Bcl-2 and the levels of GSH and FSH in ovarian granulosa cells of the overexpression SIRT1 group were increased(all P<0.05).The mRNA and protein expressions of Bax and Caspase-3 and the levels of MDA,ROS,Fe^(2+),T and LH were decreased,and the apoptosis rate of ovarian granulosa cells was decreased(all P<0.05).Conclusion:SIRT1/FOXO1 pathway is down-regulated in PCOS ovarian granu

关 键 词：多囊卵巢综合征 卵巢颗粒细胞 沉默信息调节因子1/叉头框蛋白O1信号通路 铁死亡 细胞凋亡 大鼠 

分 类 号：R36[医药卫生—病理学]