茯苓酸调节AMPK/GSK-3β/Nrf2信号通路对子宫内膜异位症大鼠内膜基质细胞铁死亡的影响  

Effect of pachymic acid on ferroptosis of endometrial stromal cells in rats with endometriosis by regulating the adenosine monophosphate-activated protein kinase/glycogen synthase kinase-3β/nuclear factor erythroid 2-related factor 2 signaling pathway

作　　者：李琳[1] 马本玲[1] 贺冰[1] 张翼[1] 邢敏 LI Lin;MA Benling;HE Bing;ZHANG Yi;XING Min(The Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410005,Hunan,China)

机构地区：[1]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《湖南中医杂志》2024年第6期175-182,共8页Hunan Journal of Traditional Chinese Medicine

基　　金：湖南省教育厅科学研究项目(21C0231)。

摘　　要：目的:探索茯苓酸调节腺苷酸活化蛋白激酶(AMPK)/糖原合成酶激酶3β(GSK-3β)/核因子E2相关因子2(Nrf2)信号通路对子宫内膜异位症(EM)大鼠基质细胞铁死亡的影响。方法:将51只SD大鼠于腹腔内异位移植子宫内膜组织构建EM大鼠模型,并将其随机分为模型组、茯苓酸低剂量(3.5 mg/kg)组、茯苓酸高剂量(7.0 mg/kg)组、茯苓酸高剂量(7.0 mg/kg)+盐酸多索啡(0.2 mg/kg)组,每组各12只,另取12只SD大鼠只开腹不移植,设为假手术组,药物分组处理后测定各组大鼠异位子宫内膜体积、异位子宫内膜组织铁死亡指标[谷胱甘肽(GSH)、丙二醛(MDA)水平、铁含量]、血清及异位子宫内膜组织炎症因子[肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)]水平、异位子宫内膜组织铁死亡相关蛋白[长链脂酰辅酶A合成酶4(ACSL4)、环加氧酶2(PTGS2)]与AMPK/GSK-3β/Nrf2通路相关蛋白表达。原代培养EM大鼠子宫内膜基质细胞,并将其随机分为对照组、茯苓酸低浓度(40 mg/L)组、茯苓酸高浓度(80 mg/L)组、茯苓酸高浓度(80 mg/L)+盐酸多索啡(10μmol/L)组,以药物分组处理后,以CCK-8法、流式细胞术分别测定各组细胞增殖及凋亡;以试剂盒检测各组细胞铁死亡指标GSH及MDA水平、铁含量;以免疫印迹实验检测各组细胞铁死亡相关蛋白及AMPK/GSK-3β/Nrf2通路相关蛋白表达。结果:与假手术组相比,模型组大鼠异位子宫内膜体积、MDA水平及铁含量、血清及异位子宫内膜组织TNF-α及IL-6水平、ACSL4及PTGS2蛋白表达升高(P<0.05),GSH水平、p-AMPK/AMPK、p-GSK-3β/GSK-3β及Nrf2蛋白表达降低(P<0.05);与模型组相比,茯苓酸低、高剂量组大鼠异位子宫内膜体积、MDA水平及铁含量、血清及异位子宫内膜组织TNF-α及IL-6水平、ACSL4及PTGS2蛋白表达降低(P<0.05),GSH水平、p-AMPK/AMPK、p-GSK-3β/GSK-3β及Nrf2蛋白表达升高(P<0.05),且高剂量茯苓酸作用更强;盐酸多索啡可减弱高剂量茯苓酸对Objective:To investigate the effect of pachymic acid on ferroptosis of endometrial stromal cells in rats with endometriosis(EM)by regulating the adenosine monophosphate-activated protein kinase(AMPK)/glycogen synthase kinase-3β(GSK-3β)/nuclear factor erythroid 2-related factor 2(Nrf2)signaling pathway.Methods:A total of 51 Sprague-Dawley rats were given ectopic transplantation of endometrial tissue in the abdominal cavity to establish a rat model of EM,and then they were randomly divided into model group,lowdose pachymic acid(3.5 mg/kg)group,high-dose pachymic acid(7.0 mg/kg)group,high-dose pachymic acid(7.0 mg/kg)+dorsomorphin dihydrochloride(0.2 mg/kg)group,with 12 rats in each group;another 12 Sprague-Dawley rats were given laparotomy without transplantation and were established as sham-operation group.Related indicators were measured after grouping and treatment,including the volume of ectopic endometrium,ferroptosis markers in ectopic endometrium(glutathione[GSH],malondialdehyde[MDA],and iron content),inflammatory factors in serum and ectopic endometrium(tumor necrosis factor-α[TNF-α]and interleukin-6[IL-6]),ferroptosis-related proteins in ectopic endometrium(acyl-CoA synthetase long-chain family member 4[ACSL4]and prostaglandin endoperoxide synthase 2[PTGS2]),and the expression of proteins associated with the AMPK/GSK-3β/Nrf2 pathway.Primary cultured endometrial stromal cells of EM rats were randomly divided into control group,low-concentration pachymic acid(40 mg/L)group,high-concentration pachymic acid(80 mg/L)group,high-concentration pachymic acid(80 mg/L)+dorsomorphin dihydrochloride(10μmol/L)group.After grouping and treatment,CCK-8 assay and flow cytometry were used to measure cell proliferation and apoptosis;kits were used to measure the levels of the ferroptosis markers GSH and MDA and the content of iron;Western blotting was used to measure the expression of ferroptosis-related proteins and proteins associated with the AMPK/GSK-3β/Nrf2 pathway.Results:Compared with the sham-operation group,th

关 键 词：茯苓酸 AMPK/GSK-3β/Nrf2 子宫内膜异位症 子宫内膜基质细胞 铁死亡 

分 类 号：R285.5[医药卫生—中药学]