摘要
目的:研究氧化苦参碱(OMT)抑制细胞外信号调节激酶ERK1/2磷酸化对醛固酮(ALD)诱导心肌细胞损伤的干预作用。方法:采用胰酶消化与差数贴壁分离纯化原代心肌细胞,免疫细胞化学分析心肌细胞纯度。实验分为空白组、模型组(1×10-5mol·L-1ALD)和OMT高(3.78×10-4mol·L-1),低剂量(1.89×10-4mol·L-1)组共4组。运用MTT法检测细胞存活率,LDH外漏法测定细胞膜损伤程度,蛋白免疫印迹法分析p-ERK1/2及ERK1/2蛋白表达,实时荧光定量PCR分析ERK mRNA表达。结果:OMT可抑制ALD诱导的细胞存活率降低和LDH外漏增加;与模型组比较,OMT预处理后p-ERK1/2蛋白表达明显下调。结论:OMT对ALD诱导的心肌细胞损伤具有保护作用,其作用与抑制ERK1/2蛋白磷酸化有关。
Objective: To observe effect of oxymatrine( OMT) on expression of extracellular signalregulated kinase 1 /2( ERK1 /2) in cardiomyocytes induced by aldosterone( ALD). Method: Pancreatic enzyme digestion method was used to extract caridac myocytes and purity of cultured caridac myocytes was evaluated by immunocytochemistry. Cardiac myocytes were pre-incubated with oxymatrine of high-does and low-does for 1 h,and then exposed to ALD for 24 h. Cell injury was evaluated by MTT assay and LDH leakage. Protein expression of ERK1 /2 and phosphorylation-ERK1 /2( p-ERK1 /2) was detected by Western blot and ERK mRNA was detected by real time polymerase chain reaction( PCR). Result: After co-incubation with OMT could enhance MTT volume and decrease LDH activity in medium induced by ALD,there was significant difference by comparing with model group. Meanwhile,expression of p-ERK1 /2 was significantly down-regulated after co-incubation with OMT.Conclusion: OMT has an protective effect on caridac myocytes induced by ALD,its mechanism may be related to inhibition of p-ERK1 /2.
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2015年第22期108-111,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81173586
81560588)
教育部新世纪人才支持计划项目(NCET-13-0747)
贵州省省长资金项目(黔科教办[2011]28号)
贵州省科技基金重点项目(黔科合JZ字[2015]2002)
贵州省高等教育科技创新团队项目(黔教合人才团队字[2014]31)
贵州省高等学校创新能力提升计划项目(黔教合协同创新字[2013]04)
贵州省高层次创新型人才项目(黔科合人才[2015]4029)
贵州省科技创新团队项目(黔科合人才团队[2015]4025号)
关键词
氧化苦参碱
心肌细胞
细胞外信号调节激酶
醛固酮
磷酸化
oxymatrine
cardiac myocytes
extracellular signal-regulated kinase
aldosterone
phosphorylation