氧化苦参碱保护醛固酮诱导的心肌细胞损伤作用机制分析

Protective Effects of Oxymatrine on Cardiomyocytes Injury Induced by Aldosteron

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.