汉黄芩苷对缺氧复氧诱导H9c2心肌细胞损伤及P38和ERK1/2表达的影响  

Effects of wogonoside on anoxia-reoxygenation induced H9c2 myocardial cells injury and expression of P38 and ERK1/2

作　　者：宋艳[1] 龚蕊 杨波[1] 王磊[1] 张玲 SONG Yan;GONG Rui;YANG Bo;WANG Lei;ZHANG Ling(Department of Critical Care Medicine,The First Affiliated Hospital of Harbin Medical University,Harbin 150001,China;Chengdu Second People's Hospital,Chengdu 610017,China)

机构地区：[1]哈尔滨医科大学第一附属医院重症医学科,黑龙江哈尔滨150001 [2]成都市第二人民医院,四川成都610017

出　　处：《西部医学》2019年第12期1820-1825,共6页Medical Journal of West China

基　　金：四川省卫生和计划生育委员会科研课题(180234)

摘　　要：目的探讨汉黄芩苷(Wog)对缺氧复氧诱导H9c2心肌细胞损伤及P38和ERK1/2表达的影响。方法培养心肌细胞H9c2构建缺氧复氧模型(A/R),给予Wog低、中、高剂量(12.5、25、50μM)处理24h,采用CCK8检测48 h后细胞活力,Hochest染色检测细胞凋亡,ELISA检测细胞上清液中心肌损伤标记物[肌酸激酶(CK),肌酸激酶同工酶(CK-MB)、肌红蛋白(Mb)]和炎性细胞因子[白介素6(IL-6)、白介素1β(IL-1β)和诱导型一氧化氮合酶(iNOS)]的含量,生化试剂盒检测氧化应激指标[超氧化物歧化酶(SOD)、丙二醛(MDA)及乳酸脱氢酶(LDH)]的含量,Western blot检测相关蛋白[B淋巴细胞瘤-2基因(Bcl-2)、Bcl-2相关x蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、Caspase-9]、丝裂原活化蛋白激酶P38及细胞外调节激酶(ERK1/2)的表达。结果低、中、高剂量Wog处理缺氧复氧模型细胞H9c2,细胞存活率、Bcl-2及SOD的表达显著升高,凋亡率、Bax、Caspase-3、Caspase-9、CK、CK-MB、Mb、IL-6、IL-1β、iNOS、MDA、LDH、P38及ERK1/2的表达显著降低(均P<0.05)。结论汉黄芩苷可通过改善炎症反应和氧化应激损伤,减少心肌细胞的凋亡,对缺氧复氧诱导H9c2心肌细胞损伤具有一定的保护作用,可能与下调P38、ERK1/2磷酸化有关。Objective To investigate the effects of wogonoside(Wog)on anoxia-reoxygenation induced H9c2 myocardial cells injury and expression of P38 and ERK1/2.Methods H9c2 myocardial cells were cultured to construct anoxia/reoxygenation(A/R)model.They were given low,medium and high doses of Wog(12.5μM,25μM,50μM)for 24 h,respectively.Cell viability after 48 h was detected by CCK8.Hochest staining was performed to detect apoptosis,ELISA was performed to detect contents ofmyocardial damage markers[creatine kinase(CK),creatine kinase isoenzyme(CK-MB),myoglobin(Mb)],and inflammatory cytokines[interleukin 6(IL-6),interleukin 1β(IL-1β)and inducible nitric oxide synthase(iNOS)]in supernate.The kits were performed to detect contents of oxidative stress indexes[superoxide dismutase(SOD),malondialdehyde(MDA)and lactate dehydrogenase(LDH)].Western blot was performed to detect the expression of apoptosis-related genes[B-cell lymphoma-2(Bcl-2),Bcl2-Associated X(Bax),Cysteinyl aspartate specific proteinase-3(caspase-3),Caspase-9],mitogen-activated protein kinase P38 and extracellular regulated protein kinases(ERK1/2).Results The low,medium and high doses of Wog was performed to process A/R model cell H9c2,the cell survival rate,Bcl-2 and expression of SOD were significantly increased,while apoptosis rate,Bax,Caspase-3,Caspase-9,CK,CK-MB,Mb,IL-6,IL-1β,iNOS,MDA,LDH,P38 and ERK1/2 expression were significantly decreased(P<0.05).Conclusion Wog can reduce cardiomyocyte apoptosis by improving inflammatory reactions and oxidative stress damage.There are certain protective effects of Wogon A/R induced H9c2 myocardial cells injury,which may be related to down-regulating phosphorylation of P38 and ERK1/2.

关 键 词：汉黄芩苷 缺氧复氧 H9C2 P38 ERK1/2 

分 类 号：R542.2[医药卫生—心血管疾病]