首页|清营生脉汤激活Akt信号通路保护H9c2心肌细胞缺氧复氧损伤的作用研究

清营生脉汤激活Akt信号通路保护H9c2心肌细胞缺氧复氧损伤的作用研究

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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.
Effect of Qingying Shengmai Decoction on Activation of Akt Signaling Pathway to Protect H9c2 Cardiomy-ocytes from Hypoxia and Reoxygenation Injury
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.

Myocardial ischemia-reperfusion injuryQingying Shengmai DecoctionSerum containing medica-tionH9c2Rats

谢蝶、付殷、赵东、张璐艳、张洋

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黑龙江中医药大学,黑龙江哈尔滨 150006

心肌缺血再灌注损伤 清营生脉汤 含药血清H9c2大鼠

黑龙江省博士后资助项目

LBH-Q19058

2024

中国中医急症
中华中医药学会

中国中医急症

CSTPCD
影响因子:1.144
ISSN:1004-745X
年,卷(期):2024.33(5)
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