芪参益气滴丸含药血清对缺氧/复氧H9C2心肌细胞KATP通道开放及PI3K/AKT信号通路影响的效应研究  

Effects of Qishen Yiqi dripping pills-containing serum on KATP channel opening and PI3K/AKT signaling pathway in hypoxic/reoxygenated H9C2 cardiocytes

作　　者：何贵新[1] 冯雨菲 秦伟彬 林琳[1] 胡梦弦 郑国坤 玉黎燕 甲子永 韦娟 文琦 HE Gui-xin;FENG Yu-fei;QIN Wei-bin;LIN Lin;HU Meng-xian;ZHENG Guo-kun;YU Li-yan;JIA Zi-yong;WEI-Juan;WEN-Qi(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530022,China;Guangxi University of Chinese Medicine,Nanning 530299,China)

机构地区：[1]广西中医药大学第一附属医院,广西南宁530022 [2]广西中医药大学,广西南宁530299

出　　处：《海南医学院学报》2022年第6期412-419,共8页Journal of Hainan Medical University

基　　金：国家自然科学基金地区基金项目(81460712);广西高校中青年教师科研基础能力提升项目(2020KY07026);广西研究生教育创新计划资助项目(YCXJ2021035)。

摘　　要：目的:探讨芪参益气滴丸含药血清对缺氧/复氧H9C2心肌细胞KATP通道开放及PI3K/AKT信号通路的影响。方法:体外培养H9C2心肌细胞随机分为5组,即A:H9C2细胞组,B:H9C2细胞+H_(2)O_(2)模型组,C:H9C2细胞+H_(2)O_(2)模型+芪参益气组,D:H9C2细胞+H_(2)O_(2)模型+芪参益气+PI3K/AKT信号通路阻断剂(wort)组,E:H9C2细胞+H_(2)O_(2)模型+芪参益气+mitoKATP特异性阻断剂(5-HD)组,按照相应的条件进行给药干预。CCK-8法检测各组心肌细胞活性;Western blot法检测各组心肌细胞AKT、pAKT蛋白的表达。标准膜片钳全细胞记录方法记录电流,由Pclamp6.0软件采集和分析电流。结果:CCK-8检测结果显示:与A组相比,B组心肌细胞活性明显下降,差异具有统计学意义(P<0.01);与B组心肌细胞相比,C组差异具有统计学意义(P<0.01);与C组相比,D、E组心肌细胞活性明显降低(P<0.05);Western blot结果显示:与A组相比,B、C、D、E组p-AKT蛋白表达显著降低,组间差异具有统计学意义(P<0.01);与B组相比,C、D、E组p-AKT蛋白表达显著升高,组间差异具有统计学意义(P<0.01);各组间AKT蛋白表达差异无统计学意义(P>0.05)。全细胞膜片钳实验结果显示:与A组相比,B组外向电流显著增加,组间差异具有统计学意义(P<0.01);与B组相比,C组的心肌细胞进一步增加外向电流,组间差异具有统计学意义(P<0.01);与C组相比,D、E组电流明显降低,组间具有统计学意义(P<0.01)。结论:芪参益气滴丸通过激活缺氧/复氧H9C2心肌细胞中p-AKT蛋白的表达及KATP通道的开放,对心肌细胞起到保护作用。Objective:To investigate the effects of Qishen Yiqi dropping pills-containing serum on KATP channel opening and PI3K/AKT signaling pathway of hypoxic/reoxygenated H9C2 cardiocytes. Methods:H9C2 cardiocytes cultured in vitro were randomly divided into five groups,A:H9C2 cell group,B:H9C2 cells + H_(2)O_(2)model group,C:H9C2 cells + H_(2)O_(2)model +Qishen Yiqi group,D:H9C2 cells + H_(2)O_(2)model + Qishen Yiqi + wort group,E:H9C2 cells + H_(2)O_(2)model + Qishenyiqi +5-HD group. The drug intervention was carried out according to the corresponding conditions. CCK-8 method was used to detect the cell activity of each group;Western blot was used to detect the expression of AKT and p-Akt proteins in myocardial cells in each group. The current was recorded by the standard patch clamp whole cell recording method,and the current was collected and analyzed by Pclamp6.0 software. Results:CCK-8 test results showed that compared with group A,the activity of myocardial cells in group B was significantly decreased,and the difference was statistically significant(P<0.01);compared with group B,the difference in group C was statistically significant(P<0.01);compared with group C,cardiomyocyte activities in groups D and E were significantly decreased,and the difference was statistically significant(P<0.05);Western blot results showed that compared with group A,p-Akt protein expression in groups B,C,D and E was significantly decreased,and the difference was statistically significant(P<0.01);compared with group B,p-Akt protein expression in groups C,D and E was significantly increased,and the difference was statistically significant(P<0.01),but there was no significant difference in AKT expression among groups(P>0.05). The results of whole cell patch clamp experiment showed that the outward current of group B was significantly increased compared with that of group A,and the difference between groups was statistically significant(P<0.01);compared with group B,cardiomyocytes in group C further increased the outward current,and the dif

关 键 词：芪参益气滴丸 H9C2 缺氧/复氧 P-AKT mitoKATP 

分 类 号：R285[医药卫生—中药学]