脉络宁注射液通过Akt信号通路降低Hcy诱导的ECV304细胞损伤的实验研究  

Effects of Mailuoning Injection on ECV304 Cell Injury Induced by Hcy through Akt Signal Pathway

作　　者：何秀芳 谭诚成 李圣永 HE Xiufang;TAN Chengcheng;LI Shengyong(Laiwu Central Hospital of Shandong Guoxin Yiyang Group,Laiwu 271103,Shandong,China)

机构地区：[1]山东国欣颐养集团莱芜中心医院,山东莱芜271103

出　　处：《中西医结合心脑血管病杂志》2022年第6期1003-1007,共5页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

摘　　要：目的探讨脉络宁注射液对同型半胱氨酸(Hcy)诱导的ECV304细胞增殖、凋亡及蛋白激酶B(Akt)信号通路、超氧化物歧化酶(SOD)活性、活性氧(ROS)、乳酸脱氢酶(LDH)、丙二醛(MDA)水平的影响。方法将ECV304细胞分为对照组、Hcy组(10 mmol/L Hcy处理细胞24 h)、Hcy+脉络宁组(10 mmol/L Hcy处理细胞24 h,加20 mg/L的脉络宁注射液处理细胞48 h)。四甲基偶氮唑盐(MTT)法检测2.5 mg/L、5 mg/L、10 mg/L、20 mg/L的脉络宁注射液对Hcy处理的ECV304细胞增殖的影响。流式细胞术检测细胞凋亡率及ROS水平。SOD试剂盒、LDH试剂盒及MDA试剂盒分别检测SOD活性及LDH和MDA水平。检测Akt、磷酸化Akt(p-Akt)、Bcl-2和Bax蛋白表达。结果5~20 mg/L的脉络宁注射液均可明显增强Hcy诱导的ECV304细胞活力。与对照组比较,Hcy组细胞凋亡率明显升高,ROS、LDH和MDA水平明显升高,SOD活性明显降低,p-Akt和Bcl-2表达明显降低,Bax表达明显升高(P<0.05)。与Hcy组比较,Hcy+脉络宁组细胞凋亡率明显降低,ROS、LDH和MDA水平明显降低,SOD活性明显升高,p-Akt和Bcl-2表达明显升高,Bax表达明显降低(P<0.05)。结论脉络宁注射液可通过降低氧化损伤、激活Akt信号通路对Hcy诱导的血管内皮细胞损伤起保护作用。Objective To investigate the effects of Mailuoning Injection on proliferation,apoptosis,protein kinase B(Akt)signaling pathway,superoxide dismutase(SOD)activity,and reactive oxygen species(ROS),lactic dehydrogenase(LDH),and malonaldehyde(MDA)levels of ECV304 cells induced by homocysteine(Hcy).Methods ECV304 cells were divided into control group,Hcy group(cells were treated with 10 mmol/L Hcy for 24 h),and Hcy+Mailuoning group(cells were treated with 10 mmol/L Hcy for 24 h and 20 mg/L Mailuoning Injection for 48 h).Methyl Thiazolyl Tetrazolium(MTT)method was used to detect the effect of 2.5 mg/L,5 mg/L,10 mg/L,20 mg/L Mailuoning on the proliferation of ECV304 cells treated with Hcy.Apoptosis rate and ROS level were detected by flow cytometry.SOD kit,LDH kit,and MDA kit were used to detect SOD activity,levels of LDH and MDA,respectively.The expression levels of Akt,phosphorylate Akt(p-Akt),Bcl-2,and Bax were detected.Results 5~20 mg/L Mailuoning Injection could significantly enhance the activity of ECV304 cells induced by Hcy.Compared with control group,the apoptosis rate in Hcy group was increased,the levels of ROS,LDH,and MDA were increased significantly,SOD activity was decreased significantly,the expression of p-Akt and Bcl-2 was decreased significantly,and Bax was increased significantly(P<0.05).Compared with Hcy group,the apoptosis rate in Hcy+Mailuoning group was decreased significantly,the levels of ROS,LDH,and MDA were decreased significantly,SOD activity was increased significantly,the expression of p-Akt and Bcl-2 was increased,Bax was decreased significantly(P<0.05).Conclusion Mailuoning Injection can protect vascular endothelial cell injury induced by Hcy to reduce oxidative damage and activating Akt signaling pathway.

关 键 词：血管内皮细胞损伤 脉络宁注射液 同型半胱氨酸 蛋白激酶B 实验研究 

分 类 号：R73[医药卫生—肿瘤]