黄芪多糖对H_2O_2所致乳鼠心肌细胞损伤保护作用的研究

Protective effects of Astraglus Polysaccharides on Neonatal Rat Cadiocytes Impared by H_2O_2

目的观察黄芪多糖(APS)对H_2O_2所致乳鼠心肌细胞损伤的保护作用机制。方法无菌条件下分离并培养乳鼠心肌细胞72 h后分为空白对照组、H_2O_2组、APS(20、40和80μmol/L)+H_2O_2组,每组设10个复孔。给药干预24 h后,观察比较各组细胞形态、细胞存活率、细胞中抗氧化酶(SOD、GSH-Px、CAT)活性和丙二醛(MDA)含量、培养液中心肌酶(AST、CPK、LDH)活性、细胞凋亡状况及细胞凋亡率情况。结果倒置光学显微镜下,空白对照组呈单层簇状生长,且伪足多而饱满,搏动明显,节律一致;H_2O_2组细胞胞浆空泡、伪足少,细胞脱落、悬浮、溶解坏死,搏动弱且频率低、节律不齐等;APS各治疗组与H_2O_2组比较,心肌细胞形态不同程度好转,且以APS(80μmol/L)+H_2O_2组改善效果最明显。H_2O_2组心肌细胞存活率与空白对照组比较降低(P<0.01);APS(40、80μmol/L)+H_2O_2组细胞存活率与H_2O_2组比较升高(P<0.01)。H_2O_2组乳鼠心肌细胞SOD、GSH-Px、CAT活性与空白对照组比较降低,MDA含量则升高(P<0.01);APS(40、80μmol/L)+H_2O_2组SOD、GSH-Px、CAT活性与H_2O_2组比较均升高(P<0.05或P<0.01),MDA含量则降低(P<0.01)。H_2O_2组乳鼠心肌细胞培养液中AST、CPK、LDH活性与空白对照组比较均升高(均P<0.01)。APS(40、80μmol/L)+H_2O_2组AST、CPK、LDH活性与H_2O_2组比较均降低(均P<0.05或P<0.01)。Flow Cytometry检测空白对照组仅存在少量凋亡细胞,H_2O_2组凋亡细胞数量较空白对照组增多;与H_2O_2干预组比较,APS干预24 h可明显改善H_2O_2诱导损伤乳鼠心肌细胞凋亡状况,其以APS(80μmol/L)+H_2O_2组效果最明显。H_2O_2组乳鼠心肌细胞凋亡率与空白对照组比较升高(P<0.01);APS(40、80μmol/L)+H_2O_2组细胞凋亡率与H_2O_2组比较则降低(P<0.01)。结论APS可通过有效改善细胞生存状态、提高细胞存活率、降低氧化应激损伤、抑制细胞凋亡而发挥对H_2O_2乳鼠心肌细胞损伤的保护作用。

Objective： To investigate the protective effects of Astraglus Polysaccharides on neonatal rat cadio- cytes impared by H2O2. Methods： Cadiocytes of neonate rat was cultivated for 72 hours and divided into six groups ： normal control group, H202 group, APS （20,40,80 μmol/L） ＋H2O2 groups （n = 10）. Twelve hours after the drugs were given,the morphology changes was observed ;the survival rate was detected by MTT;the activity of SOD, GSH-Px, CAT and the content of MDA in cardiomyocytes were determinted;the activity of AST, CPK,LDH in culture medium were detected;the cardiomyoeytes apoptosis was observed by flow cytometry and the apoptosis rate was calculated. Results： Compared with H2O2 group,the morphology of neonatal rat cadiocytes in APS groups were significantly improved;the survival rate in APS （40,80 μmo//L）＋H2O2 groups were significantly increased; the activity of SOD, GSH-Px,CAT in cardiomyocytes were significantly increased and the content of MDA were significantly decreased;the activity of AST, CPK,LDH in culture medium were significantly decreased;the ear- diomyocytes apoptosis were improved and the apoptosis rate were significantly decreased, all of the difference were significant （P〈 0.05 or P〈 0.01 ）. Conclusion： APS can effectively improve neonatal rat cadiocytes morphology, improve the activity of antioxidase and depress oxidative stress,improve the survival rate and decrease the apopto- sis rate, suggesting that APS had protective effects on neonatal rat cadiocytes impared by H2O2.