山茱萸总苷对缺氧/复氧损伤乳鼠心肌细胞钙超载的影响

Effcts of Cornus Afficinalis Glycosides on Calcium Overload of Neonatal Rats' Myocardium Cells Injuried by Hypoxia / Reoxygenation

目的:从抑制钙超载的角度探讨山茱萸总苷保护心肌细胞的作用机制。方法:采用乳鼠心肌细胞原代培养方法获得正常心肌细胞并建立缺氧/复氧损伤细胞模型,实验分正常组、模型组和山茱萸总苷低中高剂量组,培养24小时后进行细胞缺氧/复氧处理,采用Fluo4-AM荧光探针检测心肌细胞胞浆内钙离子浓度;Annexin V/PI双染法检测心肌细胞总凋亡率。结果:缺氧/复氧损伤组心肌细胞胞浆内钙离子浓度显著升高(P<0.01),山茱萸总苷低、中、高剂量可明显降低心肌组织内钙离子浓度(P<0.05,P<0.01);缺氧/复氧模型心肌细胞凋亡率明显升高,中、高剂量山茱萸总苷可明显降低心肌细胞凋亡率(P<0.01)。结论:山茱萸总苷中、高剂量预处理可以抑制缺氧/复氧损伤心肌细胞的钙超载,可能是其抗心肌细胞凋亡,保护心肌的作用机制之一。

Objective： To observe the effects of Cornus officinalis glycosides on apoptosis and[Ca^2＋]i in cultured hypoxia cardio myocytes,and explore the mechanism of myocardial protection. Methods： Primari Iy cultured neonate rats＇ cardiomyocytes were used to establish the hypoxia models,experiment was divided into normal,model and Glycosides low,middle,high doses groups. each group was cultured for 24 hours before hypoxia /reoxy genation,[Ca^2＋]i of cardiomyocytes ioaded by Fluo4-AM was analyzed by flow cytometry（ FCM）,the apoptosis percentage of cardiomyocytes was measured by Annexin V / PI FCM. Results： Calcium in hypoxia cardiac myocytes injury group was significantly higher,the intracellular calcium ion concentration of the middle and high dose Glycosides was obviously lowered,the apoptosis precentoge of cardiomyocytes in model group was obviously increased,that of middle and high dose groups was significantly decreased（ P〈0. 05）. Conclusions： Cornus officinalis glycosides can inhibit intracellular calcium overload,which may be the inhibition of cardiomyocyte apoptosis,that maybe the mechanism of myocardial protection of Cornus officinalis glycosides.