五子衍宗丸多糖对过氧化氢所致心肌细胞损伤的影响  

Effect of Polysaccharides in Wuzi Yanzong Pill on Hydrogen Peroxide-induced Cardiomyocytes Injuries

作　　者：陈桂林[1] 徐姗[1] 吴紫娟[1] 吴祎[1] CHEN Guilin;XU Shan;WU Zijuan;WU Yi(Department of Pharmacy,Changzhou Second People's Hospital,Jiangsu Province, Changzhou 213000,China)

机构地区：[1]江苏省常州市第二人民医院药学部,常州213000

出　　处：《医药导报》2021年第3期306-310,共5页Herald of Medicine

基　　金：江苏省天晴医院药学基金资助项目(Q2018105);2018年常州市科学技术局第二十六批科技计划(应用基础研究指导性)项目(CJ20189007)。

摘　　要：目的研究五子衍宗丸多糖对过氧化氢(H_(2)O_(2))诱导心肌细胞损伤的影响及其作用机制。方法将小鼠心肌细胞随机分为阴性对照组(正常小鼠心肌细胞分离纯化后培养)、诱导组(心肌细胞培养液加入100μmol·L^(-1) H_(2)O_(2)反应2 h)、五子衍宗丸多糖大剂量组(1×10^(-4) mol·L^(-1)五子衍宗丸多糖培养24 h,100μmol·L^(-1)H_(2)O_(2)反应2 h)、五子衍宗丸多糖中剂量组(5×10^(-5)mol·L^(-1)五子衍宗丸多糖培养24 h,100μmol·L^(-1)H_(2)O_(2)反应2 h)、五子衍宗丸多糖小剂量组(1×10^(-5) mol·L^(-1)五子衍宗丸多糖培养24 h,100μmol·L^(-1)H_(2)O_(2)反应2 h)。噻唑蓝(MTT)比色法检测细胞乳酸脱氢酶(LDH)、肌酸磷酸激酶(CK)、丙二醛(MDA)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)及谷胱甘肽过氧化物酶(GSH-Px)含量;酶联免疫吸附(ELISA)法检测白细胞介素-6(IL-6)及IL^(-1)2;免疫蛋白印迹法检测B淋巴细胞瘤-2基因(Bcl-2)、细胞凋亡调节因子Bax和含半胱氨酸的天冬氨酸蛋白水解酶(caspase)蛋白。结果与阴性对照组比较,诱导组心肌细胞吸光度(A值)降低,LDH和CK、MDA、IL-6与IL^(-1)2释放量升高,CAT(五子衍宗丸多糖大剂量组)、SOD和GSH-Px(五子衍宗丸多糖中、大剂量组)活性降低,差异有统计学意义(P<0.05);与诱导组比较,五子衍宗丸多糖各剂量组A值、CAT(五子衍宗丸多糖大剂量组)、SOD和GSH-Px五子衍宗丸多糖中、大剂量组升高;IL-6、IL^(-1)2、LDH、CK和MDA(五子衍宗丸多糖大剂量组)释放量降低;Bcl-2和caspase蛋白表达下调,Bax蛋白表达上调,均差异有统计学意义(P<0.05)。结论五子衍宗丸多糖能有效缓解心肌细胞过氧化损伤,减轻或抑制脂质过氧化反应,缓解炎症因子对心肌细胞的损伤,作用机制可能与下调Bcl-2及caspase蛋白表达、上调Bax蛋白表达有关。Objective To study the effect and mechanism of polysaccharides in Wuzi Yanzong pill on cardiomyocytes injuries induced by hydrogen peroxide(H_(2)O_(2)).Methods Cultured mouse cardiomyocytes were used to establish a model of H_(2)O_(2)-induced cardiomyocyte injury.Cultured cells were randomly divided into five groups:negative control group,induction group,high dose group,middle dose group,and low dose group.In negative control group,normal mouse cardiomyocytes were isolated and purified and cultured;In the induction group,100μmol·L^(-1) H_(2)O_(2) was added into cardiomyocyte culture medium for 2 hours;In the high dose group,Polysaccharide Wuzi Yanzong was added and treated for 24 hours at a concentration of 1×10^(-4) mol·L^(-1),and 100μmol·L^(-1) H_(2)O_(2) was added for 2 hours.In the middle dose group,the cardiomyocytes were cultured at the concentration of 5×10^(-5 )mol·L^(-1) for 24 hours and then reacted with 100μmol·L^(-1) H_(2)O_(2) for 2 hours.In the low dose group,the cardiomyocytes were cultured at the concentration of 1×10^(-5) mol·L^(-1) for 24 hours and reacted with 100μmol·L^(-1) H_(2)O_(2) for 2 hours.The protective effect of polysaccharide in Wuzi Yanzong on cardiomyocytes was detected by MTT method.The cell lactate dehydrogenase(LDH)and creatine phosphokinase(CK)were detected by kit.The contents of CK,MDA,CAT,SOD and GSH-Px were detected,the changes of IL-6 and IL^(-1)2 were detected by ELISA kit,and the expressions of Bcl-2,Bax and Caspase were detected by Western blot.Results Compared with the negative control group,the absorbance(A value)of cardiomyocytes in the induction group was decreased in MTT cytes,the release of LDH and CK,MDA,IL-6 and IL^(-1)2 was increased,and the activities of SOD,GSH-Px and CAT were decreased in the induction group.The difference was statistically significant(P<0.05).Compared with the induction group,the A value,and the activities of SOD,CAT and GSH-Px were increased;the release of IL-6,IL^(-1)2,LDH,CK and MDA was decreased;the protein expression of Bc

关 键 词：五子衍宗丸多糖 过氧化氢 心肌细胞损伤 

分 类 号：R286[医药卫生—中药学] R965[医药卫生—中医学]