Ang1-7、A779、AngⅡ对胰岛素抵抗的大鼠卵巢颗粒细胞胰岛素信号通路蛋白的影响  

Effects of Ang1-7,A779 and AngⅡon insulin signaling pathway proteins in ovarian granulosa cells of insulin resistant rat

作　　者：陈思 王秀丽[2] 何小静[1] 张媛[1] 王怡倩 李晓冬[1] Chen Si;Wang Xiuli;He Xiaojing;Zhang Yuan;Wang Yiqian;Li Xiaodong(Department of Gynecology,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China;Department of Clinical Laboratory,the Second Hospital of Hebei Medical University,Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第二医院妇科,石家庄050000 [2]河北医科大学第二医院检验科,石家庄050000

出　　处：《中华生殖与避孕杂志》2021年第2期131-136,共6页Chinese Journal of Reproduction and Contraception

基　　金：河北医科大学第二医院院基金(2h201822)。

摘　　要：目的探讨Ang1-7、Mas受体拮抗剂A779、AngⅡ作用后,对胰岛素抵抗的大鼠卵巢颗粒细胞葡萄糖摄取及胰岛素信号通路蛋白的影响。方法①构建大鼠卵巢颗粒细胞胰岛素抵抗模型:将颗粒细胞分为空白组和模型组,模型组细胞用地塞米松、胰岛素进行处理,检测两组细胞培养基上清液24 h内葡萄糖浓度、乳酸浓度的差值。②进一步将模型组细胞分别加入不同药物处理24 h:AngⅡ组、Ang1-7组、Ang1-7+AngⅡ组、A779组、Ang1-7+A779组,并检测药物作用后细胞培养基上清液24 h内葡萄糖浓度差值。③采用Western blotting检测空白组、模型组、各药物处理组颗粒细胞Akt、GSK-3β、AS160及其磷酸化蛋白(P-Akt、P-GSK-3β、P-AS160)以及Mas受体蛋白的表达。结果①跟空白组相比,模型组的葡萄糖浓度差值小、乳酸浓度差值小,提示颗粒细胞胰岛素抵抗模型建立成功。②与模型组比较,Ang1-7组葡萄糖浓度差值大(5.55±0.21,P=0.0021),AngⅡ组差值小,Ang1-7+AngⅡ组、A779组差异无统计学意义(P>0.05),Ang1-7+A779组葡萄糖浓度差值小(4.89±0.20,P=0.0108)。③空白组、模型组、各药物处理组颗粒细胞的Akt、GSK-3β、AS160的表达无明显差异。与模型组比较,Ang1-7组P-Akt、P-GSK-3β、P-AS160的表达增强,AngⅡ组P-Akt、P-GSK-3β、P-AS160的表达减弱。与Ang1-7组比较,Ang1-7+A779组Mas受体表达量降低。结论①Ang1-7作用后,胰岛素抵抗的大鼠卵巢颗粒细胞葡萄糖浓度差值增加,AngⅡ作用后与Ang1-7相反,二者相互拮抗,同时Ang1-7、AngⅡ作用后颗粒细胞胰岛素信号通路关键蛋白P-Akt、P-GSK-3β及P-AS160也出现相反变化,表明通过细胞葡萄糖浓度差值体现的细胞葡萄糖代谢有可能是胰岛素信号通路蛋白P-Akt、P-GSK-3β及P-AS160表达变化的结果。②Ang1-7作用后受体Mas表达上调,加入Mas受体拮抗剂A779,细胞葡萄糖代谢受抑制,P-Akt、P-GSK-3β及P-AS160表达降低,提示Ang1-7改善Objective To determine the changes of glucose and insulin signaling pathway proteins in rat ovarian insulin resistant granulosa cells by detecting Ang1-7,MAS receptor inhibitors A779 and AngⅡ.Methods First,rat ovarian granulosa cells were cultured with dexamethasone and insulin to establish the insulin resistance(IR)model,and the concentration differences of glucose and lactic acid in both blank and IR model groups were detected.According to the different drugs treated,the cells in the IR model group were divided into AngⅡgroup,Ang1-7 group,Ang1-7+AngⅡgroup,A779 group,Ang1-7+A779 group.After 24 h,the concentration differences of glucose of all groups were tested.At last,Western blotting was used to detect the expressions of P-Akt/Akt,P-Gsk-3β/GSK-3β,P-AS160/AS160 in the Mas receptor protein and key proteins in the insulin signaling pathway in the above groups.Results 1)Compared with the blank group,the concentration differences of glucose and lactic acid of the supernatant were reduced after applying dexamethasone and insulin to rat ovarian granulosa cells,suggesting that the IR model was successfully established.2)Compared with the model group,the concentration difference of glucose in the Ang1-7 group was increased(5.55±0.21,P=0.0021),while the AngⅡsubgroup was decreased.There was no significant difference of the concentration difference of glucose between the Ang1-7+AngⅡgroup and the A779 group(P>0.05).However,the concentration difference of glucose in the Ang1-7+A779 group was decreased(4.89±0.20,P=0.0108).3)There was no difference in the expressions of Akt,GSK-3β,AS160 among the black group,model group and the different drugs treated groups.Compared with the model group,the expressions of P-Akt,P-GSK-3β,and P-AS160 in the Ang1-7 group were enhanced,while the expressions of P-Akt,P-GSK-3β,and P-AS160 in the AngⅡgroup were decreased.And compared with Ang1-7 group,Mas receptor expression in Ang1-7+A779 group was decreased.Conclusion 1)The effect of Ang1-7 on improving the IR condition of rat

关 键 词：多囊卵巢综合征 胰岛素抵抗 血管紧张素Ⅱ 

分 类 号：R711.75[医药卫生—妇产科学]