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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
宋艳,龚蕊,杨波,王磊,张玲
0
(哈尔滨医科大学第一附属医院重症医学科;成都市第二人民医院)
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、50 μM)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。
关键词:  汉黄芩苷  缺氧复氧  H9c2  P38  ERK1/2
DOI:
基金项目:四川省卫生和计划生育委员会科研课题(180234)
Effects of wogonoside on anoxia reoxygenation induced H9c2 myocardial cells injury and expression of P38 and ERK1/2
SONG Yan,GONG Rui,YANG Bo,WANG Lei,ZHANG Ling
(Department of Critical Care Medicine, The First Affiliated Hospital of Harbin Medical University;Chengdu Second People′s Hospital)
Abstract:
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, 50 μM) for 24h, respectively. Cell viability after 48h 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 [Bcell lymphoma2(Bcl-2),Bcl2Associated 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, I-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.
Key words:  Wogonoside  Anoxia reoxygenation  H9c2  P38  ERK1/2

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