黄连温胆汤通过PI3K/Akt通路对缺氧复氧诱导心肌细胞损伤作用机制  

Impacts and mechanism of Huanglian Wendan decoction on myocardial cell injury induced by hypoxia reoxygenation through PI3K/Akt pathway

作　　者：王琪[1] 吴限[1] 刘丰[1] 苗永悦 孙少谦 刘莉[1] WANG Qi;WU Xian;LIU Feng;MIAO Yongyue;SUN Shaoqian;LIU Li(First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学附属第一医院,黑龙江哈尔滨150040

出　　处：《陕西中医》2023年第7期859-863,共5页Shaanxi Journal of Traditional Chinese Medicine

基　　金：黑龙江省中医药管理局科研项目(ZHY2023-183);第七批全国老中医药专家学术经验继承工作项目[国中医药人教函(2022)76号]。

摘　　要：目的:探讨黄连温胆汤通过磷脂酰肌醇3激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(Akt)通路对缺氧复氧诱导心肌细胞损伤的作用机制。方法:构建缺氧复氧诱导的大鼠心肌细胞H9C2损伤模型,分为对照组、缺氧复氧组、黄连温胆汤低剂量组、黄连温胆汤中剂量组、黄连温胆汤高剂量组。CCK-8法检测H9C2细胞增殖。流式细胞术检测H9C2细胞凋亡。检测细胞内活性氧(ROS)、细胞上清中超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量。Western blotting检测细胞中Bcl-2相关X蛋白(Bax)、B淋巴细胞瘤-2(Bcl-2)、p-PI3K、p-Akt蛋白表达。结果:缺氧复氧组H9C2细胞OD 450值、SOD活性、Bcl-2、p-PI3K、p-Akt蛋白表达均低于对照组,而细胞凋亡率、ROS平均荧光强度、MDA含量、Bax蛋白表达高于对照组,差异有统计学意义(均P<0.05)。黄连温胆汤低剂量组、黄连温胆汤中剂量组、黄连温胆汤高剂量组H9C2细胞OD 450值、SOD活性、Bcl-2、p-PI3K、p-Akt蛋白表达高于缺氧复氧组,而细胞凋亡率、ROS平均荧光强度、MDA含量、Bax蛋白表达低于缺氧复氧组,差异有统计学意义(均P<0.05)。结论:黄连温胆汤通过激活PI3K/Akt信号通路减轻缺氧复氧诱导的H9C2细胞损伤。Objective:To investigate impact and mechanism of Huanglian Wendan decoction on myocardial cell injury induced by hypoxia reoxygenation through phosphatidylinositol-3-kinase(PI3K)/serine threonine protein kinase(Akt)pathway.Methods:A model of H9C2 injury in rat cardiomyocytes induced by hypoxia reoxygenation was established and grouped into control group,hypoxia reoxygenation group,low dose Huanglian Wendan decoction group,medium dose Huanglian Wendan decoction group,high dose Huanglian Wendan decoction group.CCK-8 method was used to detect proliferation of H9C2 cells.The apoptosis of H9C2 cells was detected by flow cytometry.Intracellular reactive oxygen species(ROS),superoxide dismutase(SOD)activity and malondialdehyde(MDA)content in cell supernatant were detected.Western blotting was applied to detect expression of Bcl-2 associated X protein(Bax),B cell-lymphomatoma-2(Bcl-2),p-PI3K,and p-Akt proteins in the cells.Results:The OD 450 value,SOD activity,protein expression of Bcl-2,p-PI3K and p-Akt in H9C2 cells in the hypoxia reoxygenation group were lower than those in the control group,and apoptosis rate,ROS average fluorescence intensity,content of MDA,and protein expression of Bax were higher,difference statistically significant(all P<0.05).The OD 450 value,SOD activity,protein expression of Bcl-2,p-PI3K and p-Akt in H9C2 cells in the low,medium,and high dose Huanglian Wendan decoction groups were higher than those in the hypoxia reoxygenation group,while the apoptosis rate,ROS average fluorescence intensity,content of MDA,and protein expression of Bax lower,difference statistically significant(all P<0.05).Conclusion:Huanglian Wendan decoction may alleviate injury of H9C2 cells induced by hypoxia reoxygenation by activating the PI3K/Akt signaling pathway.

关 键 词：黄连温胆汤 磷脂酰肌醇3激酶/丝氨酸苏氨酸蛋白激酶通路 缺氧复氧 心肌损伤 凋亡 氧化应激 

分 类 号：R289.5[医药卫生—方剂学]