摘要
目的研究清营生脉汤含药血清对缺氧复氧诱导H9c2心肌细胞损伤的保护机制。方法本研究采用H9c2心肌细胞缺氧/复氧(H/R)方法模拟心肌缺血再灌注损伤(MIRI)细胞模型,细胞实验分组为:空白血清对照组、模型组、含药血清低剂量组、含药血清中剂量组、含药血清高剂量组。应用CCK-8法测定心肌细胞活性,流式细胞术检测细胞凋亡率,ELISA法检测超氧化物歧化酶(SOD)、乳酸脱氢酶(LDH)、丙二醛(MDA)、肌酸激酶同工酶(CK-MB)水平,Western blotting法检测细胞内磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(Akt)及p-Akt的蛋白表达。结果与对照组相比,模型组细胞存活率显著降低、凋亡率显著升高、细胞内SOD水平显著降低,LDH、MDA、CK-MB水平显著升高(P<0.01);与模型组相比,含药血清低、中、高剂量组细胞的存活率显著升高、凋亡率显著降低、细胞内SOD水平显著升高,LDH、MDA、CK-MB水平显著降低(P<0.01);与对照组相比,模型组细胞中p-Akt蛋白表达水平显著降低(P<0.01),p-Akt/Akt比值降低(P<0.05);与模型组相比,含药血清高剂量组细胞内p-Akt蛋白表达量显著升高(P<0.01),p-Akt/Akt比值明显升高(P<0.05)。结论清营生脉汤含药血清能够提高缺氧复氧心肌细胞的活性,降低细胞凋亡率,升高SOD水平,降低LDH、MDA、CK-MB水平,提高细胞抗氧化能力,且可能通过促进Akt磷酸化降低缺氧复氧诱导的H9c2心肌细胞损伤程度。
Objective:To study the protective mechanism of serum containing Qingying Shengmai Decoction on H9c2 cardiomyocyte injury induced by hypoxia and reoxygenation.Methods:In this study,H9c2 cardiomyocyte hypoxia/reoxygenation(H/R)method was used to simulate the cell model of myocardial ischemia-reperfusion injury(MIRI).The cell experiment was divided into blank serum control group,model group,low-dose drug-containing serum group,medium-dose drug-containing serum group,and high-dose drug-containing serum group.Cardiomyo⁃cyte activity was determined by CCK-8 method,apoptosis rate was detected by flow cytometry,SOD,LDH,MDA and CK-MB levels were detected by ELISA,and PI3K,Akt and p-Akt protein expression were detected by West⁃ern blotting method.Results:Compared with blank group,the cell survival rate in model group was significantly decreased,the apoptosis rate was significantly increased,the SOD level was significantly decreased,and the levels of LDH,MDA and CK-MB were significantly increased(P<0.01).Compared with model group,the survival rate was significantly increased,the apoptosis rate was significantly decreased,the SOD level was significantly in⁃creased,and the levels of LDH,MDA and CK-MB were significantly decreased in low,medium and high dose groups(P<0.01).Compared with control group,p-Akt protein expression level were significantly decreased(P<0.01)and p-Akt/Akt ratio were decreased in model group(P<0.05);compared with model group,intracellular p-Akt protein expression were significantly increased(P<0.01)and p-Akt/Akt ratio were increased in drug-contain⁃ing serum high-dose group(P<0.05).Conclusion:Serum containing Qingying Shengmai Decoction can improve the activity of hypoxic and reoxygenated cardiomyocytes,reduce the apoptosis rate,increase SOD level,decrease LDH,MDA and CK-MB levels,improve the antioxidant capacity of cells,and may reduce the damage degree of H9c2 cardiomyocytes induced by hypoxic and reoxygenation by promoting Akt phosphorylation.
作者
谢蝶
付殷
赵东
张璐艳
张洋
Xie Die;Fu Yin;Zhao Dong;Zhang Luyan;Zhang Yang(Heilongjiang University of Chinese Medicine,Heilongjiang,Harbin 150006,China)
出处
《中国中医急症》
2024年第5期804-808,共5页
Journal of Emergency in Traditional Chinese Medicine
基金
黑龙江省博士后资助项目(LBH-Q19058)。
关键词
心肌缺血再灌注损伤
清营生脉汤
含药血清
H9C2
大鼠
Myocardial ischemia-reperfusion injury
Qingying Shengmai Decoction
Serum containing medica⁃tion
H9c2
Rats
