丹参多酚酸盐对缺氧心肌细胞的保护作用

Protective effect of Salvianolate on cardiomyocytes under hypoxia

目的分析丹参多酚酸盐对缺氧心肌细胞的保护作用。方法新生SD大鼠50只,分离并培养其原代心肌细胞,选择生长良好的心肌细胞,建立心肌细胞缺氧/复氧(hypoxia/reoxygenation,H/R)模型,随机分为空白对照组(仅于细胞DMEM培养基中培养)、缺氧/复氧组(仅建立H/R模型)、干预1组(H/R模型+丹参多酚酸盐)、干预2组(丹参多酚酸盐+H/R模型)、干预3组(丹参多酚酸盐+H/R模型+丹参多酚酸盐)各10只,采用四甲基偶氮唑盐比色法(MTT)测定心肌细胞活力,以紫外分光光度计平行测定各组氧化应激指标、心肌损伤标志物、炎症因子水平。结果缺氧/复氧组心肌细胞存活率低于空白对照组,干预1组、干预2组、干预3组细胞存活率高于缺氧/复氧组,且药物干预组中干预3组细胞存活率最高(P<0.05);缺氧/复氧组SOD含量低于空白对照组,而MDA、p22phox活性明显高于空白对照组,干预1组、干预2组、干预3组SOD含量高于缺氧/复氧组,而MDA、p22phox活性低于缺氧/复氧组,且干预3组上述指标改善最明显(P<0.05);缺氧/复氧组心肌细胞培养液中AST、CK⁃MB、LDH、TNF⁃α、IL⁃1、hs⁃CRP水平明显高于空白对照组,而干预1组、干预2组、干预3组AST、CK⁃MB、LDH、TNF⁃α、IL⁃1、hs⁃CRP水平明显低于缺氧/复氧组,且干预3组上述指标改善最明显(P<0.05)。结论丹参多酚酸盐可较好保护缺氧心肌细胞,减轻其氧化应激反应、心肌损伤、炎症因子水平,抑制细胞凋亡,提高细胞存活率。

Objective To determine the protective effect of Salvianolate on cardiomyocytes under hypoxia.Methods Fifty neonatal SD rats were included and their primary cardiomyocytes were isolated and cultured.The well⁃developed cardiomyocytes were selected to establish the hypoxia/reoxygenation(H/R)models of cardiomyocytes.The rats were randomly divided into the blank control group(cultured in cell DMEM medium alone),H/R group(only establishing H/R model),intervention group 1(H/R model+Salvianolate),intervention group 2(Salvianolate+H/R model)and intervention group 3(Salvianolate+H/R model+Salvianolate),with n=10 rats in each group.The viability of cardiomyocytes was determined by methyl thiazolyl tetrazolium(MTT)assay.The levels of oxidative stress indicators,myocardial injury markers and inflammatory factors were determined by ultraviolet spectrophotometer.Results The survival rate of cardiomyocytes in the H/R group was lower than that in the blank control group.The survival rate of cardiomyocytes in the intervention groups 1,2 and 3 was higher than that in the H/R group,respectively.The survival rate was the highest in the intervention group 3 among the three intervention groups(P<0.05).The level of SOD in the H/R group was lower than that in the blank control group,whereas the MDA and p22phox activities were significantly higher than those in the blank control group.The level of SOD in the intervention groups 1,2 and 3 was higher than that in the H/R group,respectively,whereas the MDA and p22phox activities were lower than those in the H/R group.The improvement of the indicators was the most obvious in the intervention group 3(P<0.05).The levels of AST,CK⁃MB,LDH,TNF⁃α,IL⁃1 and hs⁃CRP in the culture medium of cardiomyocytes in the H/R group were significantly higher than those in the blank control group.The levels of AST,CK⁃MB,LDH,TNF⁃α,IL⁃1 and hs⁃CRP in the intervention groups 1,2 and 3 were significantly lower than those in the H/R group,and the improvement of the indicators was the most obvious in the intervention group 3(P<0.05).Conclusion Salvianolate may well protect cardiomyocytes under hypoxia,relieve oxidative stress and myocardial damage,decrease levels of inflammatory factors,inhibit cell apoptosis,and increase the survival rate of cells.