[1]白 俊,赵晓鹏,王海静,等.补肾活血法调控PI3K/AKT/mTOR通路修复卵巢颗粒细胞自噬损伤机制研究[J].陕西中医,2023,(5):547-551.[doi:DOI:10.3969/j.issn.1000-7369.2023.05.001]
　BAI Jun,ZHAO Xiaopeng,WANG Haijing,et al.Research on the mechanism of repairing autophagic damage in ovarian granulosa cells by regulating PI3K/AKT/mTOR pathway by tonifying kidney and activating blood[J].,2023,(5):547-551.[doi:DOI:10.3969/j.issn.1000-7369.2023.05.001]
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补肾活血法调控PI3K/AKT/mTOR通路修复卵巢颗粒细胞自噬损伤机制研究
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2023年5期
页码:
547-551
栏目:
基础研究
出版日期:
2023-05-05

文章信息/Info

Title:
Research on the mechanism of repairing autophagic damage in ovarian granulosa cells by regulating PI3K/AKT/mTOR pathway by tonifying kidney and activating blood
作者:
白 俊1; 赵晓鹏1; 王海静1; 王丽君1; 张 辉1; 李 楠1; 陈 梅1; 贺丰杰1; 吴克明2
(1.陕西中医药大学附属医院,陕西 咸阳 712000; 2.成都中医药大学附属医院,四川 成都 610075)
Author(s):
BAI Jun; ZHAO Xiaopeng; WANG Haijing; WANG Lijun; ZHANG Hui; LI Nan; CHEN Mei; HE Fengjie; WU Keming
(Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712000,China)
关键词:
补肾活血法;  卵巢颗粒细胞;  自噬;  雷公藤;  PI3K/AKT/mTOR通路;  卵泡发育障碍
Keywords:
Tonifying kidney and activating blood;  Ovarian granulosa cells;  Autophagy;  Tripterygium wilfordii;  PI3K/AKT/mTOR signaling pathway;  Follicular dysplasia
分类号:
R 711.75
DOI:
DOI:10.3969/j.issn.1000-7369.2023.05.001
文献标志码:
A
摘要:
目的:观察补肾活血法调控磷脂酰肌醇3-激酶/蛋白激酶B/哺乳动物雷帕霉素靶蛋白(PI3K/AKT/mTOR)通路对体外培养大鼠卵巢颗粒细胞(OGCs)自噬损伤的影响。方法:将原代培养的大鼠卵巢颗粒细胞分为六组:空白对照组、模型组、中药低剂量组、中药中剂量组、中药高剂量组、果纳芬组。采用ELISA法检测各组中雌二醇(E2)、孕酮(P)分泌量; Western blot法和免疫荧光法分别检测各组OGCs中PI3K、AKT、mTOR及自噬关键分子酵母Atg6 同系物1(Beclin 1)、微管相关蛋1轻链3(LC3)、自噬受体蛋白p62的蛋白表达。结果:模型组E2含量显著降低,中药各剂量组E2含量均上升(均P<0.05),且中药高剂量组与果纳芬组E2含量相当; 各组对P的含量均没有影响(均P>0.05)。中药各剂量组及果纳芬组p-PI3K、p-AKT、p-mTOR水平和p62的表达上调,Beclin 1和LC3Ⅱ/LC3Ⅰ的表达下调,且中药高剂量组效果优于果纳芬组(均P<0.05)。结论:补肾活血中药复方新加归肾丸可能是通过激活PI3K/AKT/mTOR通路,下调下游自噬效应分子Beclin 1及LC3的表达并减少p62的消耗来修复OGCs的自噬损伤。
Abstract:
Objective:To observe the effect of PI3K/AKT/mTOR pathway regulation by kidney tonic and blood activation method on autophagic injury in cultured rat ovarian granulosa cells in vitro.Methods:Primary cultured rat ovarian granulosa cells were divided into 6 groups:blank control group,model group,Chinese medicine low-dose group,Chinese medicine medium-dose group,Chinese medicine high-dose group and Gonal-F group.The E2 and P content and the expression of PI3K,AKT,mTOR,Beclin 1,LC3 and p62 proteins in each group of OGCs.were detected by WB method and immunofluorescence method,respectively.Results:E2content were detected by was significantly decreased in the model group and increased in all dose groups of Chinese medicine(all P<0.05),and the E2content in the high dose group of Chinese medicine was comparable to that in the Gonal-F group.There was no effect of each group and on the content of P(P>0.05).The expression of p-PI3K,p-AKT,p-mTOR levels and p62 were up-regulated in each dose group of Chinese medicine and Gonal-F group,and the expression of Beclin 1 and LC3Ⅱ/LC3Ⅰ were down-regulated,and the effect of high-dose group of Chinese medicine was better than that of western medicine Gonal-F group(all P<0.05).Conclusion:The results suggest that the kidney tonic and blood activating Chinese herbal medicine compound Xinjia Guishen Wan may repair autophagic damage in ovarian granulosa cells by activating the PI3K/AKT/mTOR pathway,down-regulating the expression of downstream autophagic effector molecules Beclin 1 and LC3 and reducing the depletion of p62.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(81873334); 陕西省自然科学基础研究项目(2023-JC-QN-0899); 陕西省教育厅专项科研计划项目(20JK0596); 陕西省咸阳市科技计划项目(2021ZDYF-SF-0033); 陕西中医药大学附属医院院级课题(2020QN002); 全国名老中医药专家传承工作室建设项目
更新日期/Last Update: 2023-05-09