摘要
目的:观察葡萄籽原花青素对多柔比星(DOX)诱导的心脏毒性以及核因子 E2 相关基因(Nrf2)/血红素加氧酶-1(HO-1)通路的作用.方法:通过 DOX构建小鼠急性心脏毒性模型,本实验分为对照组(正常小鼠)、葡萄籽原花青素组、DOX组、DOX+葡萄籽原花青素组、DOX+葡萄籽原花青素+ML385组.超声心动图检查小鼠心功能;苏木精-伊红(HE)染色观察小鼠心肌病理改变;原位末端标记测定法(TUNEL)检测小鼠心肌细胞凋亡情况;酶联免疫吸附法(ELISA)测定血清肌酸激酶同工酶(CK-MB)、白细胞介素-6(IL-6)、白细胞介素-1β(IL-1β)水平;试剂盒检测心肌组织中活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)水平;蛋白质免疫印迹法(Western Blot)测定 Nrf2/HO-1通路蛋白表达水平.结果:与对照组比较,DOX组小鼠心肌损伤加重,左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD),血清 CK-MB、IL-6、IL-1β水平及心肌组织 ROS、MDA及心肌细胞凋亡率升高(P<0.05),射血分数(EF)、短轴缩短分数(FS)、心肌组织 SOD、Nrf2、HO-1 蛋白水平降低(P<0.05);葡萄籽原花青素组小鼠心肌损伤,LVESD、LVEDD、EF、FS水平,血清 CK-MB、IL-6、IL-1β水平,心肌组织 ROS、MDA、SOD、Nrf2、HO-1 蛋白,心肌细胞凋亡率差异均无统计学意义(P>0.05).与 DOX组比较,DOX+葡萄籽原花青组小鼠心肌损伤减轻,LVESD、LVEDD降低,血清 CK-MB、IL-6、IL-1β水平降低,心肌组织 ROS、MDA、心肌细胞凋亡率降低(P<0.05),EF、FS及心肌组织 SOD、Nrf2、HO-1蛋白水平升高(P<0.05);与DOX+葡萄籽原花青素组比较,DOX+葡萄籽原花青素+ML385 组小鼠心肌损伤加重,LVESD、LVEDD及血清 CK-MB、IL-6、IL-1β水平、心肌组织 ROS、MDA、心肌细胞凋亡率升高(P<0.05),EF、FS、心肌组织SOD、Nrf2、HO-1蛋白水平降低(P<0.05).结论:葡萄籽原花青素可能通过激活 Nrf2/HO-1通路,对 DOX诱导的心脏毒性小鼠发挥保护作用.
Abstract
Objective:To observe grape seed proanthocyanidins could alleviate doxorubicin(DOX)-induced cardiotoxicity and the role of nuclear factor E2-related gene(Nrf2)/heme oxygenase-1(HO-1)pathway on it.Methods:The mice model with acute cardiotoxicity was established by DOX,and the experiment was divided into the control group(normal mice),grape seed proanthocyanidin group,DOX group,DOX+grape seed proanthocyanidin group,DOX+grape seed proanthocyanidin+ML385 group.Echocardiography was used to detect cardiac function in mice;hematoxylin-eosin(HE)staining was used to observe the pathological changes of myocardial cells in mice;the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)assay was used to detect the apoptosis of cardiomyocytes;enzyme-linked immunosorbent assay(ELISA)was used to measure serum creatine kinase isoenzyme(CK-MB),interleukin-6(IL-6),and interleukin-1β(IL-1β)levels;The kit was used to detect reactive oxygen species(ROS),malondialdehyde(MDA),and superoxide dismutase(SOD)levels in myocardial tissue;Western Blot was used to determine the expression levels of Nrf2/HO-1 pathway proteins.Results:Compared with the control group,the myocardial injury of the mice in the DOX group aggravated;the left ventricular end-diastolic diameter(LVEDD),left ventricular end-systolic diameter(LVESD),serum CK-MB,IL-6,IL-1β levels,myocardial tissue ROS,MDA,myocardial cell apoptosis rate increased(P<0.05);ejection fraction(EF),fractional shortening(FS),myocardial tissue SOD,Nrf2,and HO-1 protein levels decreased(P<0.05).In the seed proanthocyanidin group,the difference in myocardial injury,LVESD,LVEDD,EF,FS,serum CK-MB,IL-6,IL-1βlevels,myocardial tissue ROS,MDA,SOD,Nrf2,HO-1 protein,and cardiomyocyte apoptosis rate was not statistically significant(P>0.05).Compared with the DOX group,the myocardial injury of the mice in the DOX+grape seed proanthocyanidin group alleviated;LVESD,LVEDD,serum CK-MB,IL-6,IL-1β levels,myocardial tissue ROS,MDA,and myocardial cell apoptosis rate reduced(P<0.05);EF,FS,protein levels of SOD,Nrf2,and HO-1 in myocardial tissue increased(P<0.05).Compared with the DOX+ grape seed proanthocyanidin group,myocardial injury in the DOX+ grape seed proanthocyanidin+ ML385 group aggravated;LVESD,LVEDD,Serum CK-MB,IL-6,IL-1βlevels,myocardial tissue ROS,MDA,and myocardial cell apoptosis rate increased(P<0.05);EF,FS,myocardial tissue SOD,Nrf2,and HO-1 protein levels decreased(P<0.05).Conclusion:Grape seed proanthocyanidins may protect against DOX-induced cardiotoxicity in mice by activating the Nrf2/HO-1 pathway.
基金项目
天津市科技发展计划项目(2018-HBCD-18)
维普
万方数据