基于氧化应激探讨稳心丹对心肌缺血再灌注损伤大鼠心肌保护作用机制  

Discussion on the protective effect and mechanism of Wenxin Dan on myocardial ischemia-reperfusion injury rats based on oxidative stress

作　　者：王秀珍[1,2,3] 邹伟[1,4] 李冀[1] 沈晓璞[1,2] 王特哈斯[1] 彭宇飞 WANG Xiu-zhen;ZOU Wei;LI Ji;SHEN Xiao-pu;WANG Te-hasi;PENG Yu-fei(Heilongjiang University of Chinese Medicine,Harbin 150040,China;The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150001,China;Hanan Branch,The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine,Harbin 150000,China;First Affiliated Hospital,Heilongjiang University of Chinese Medicine,Harbin 150040,China)

机构地区：[1]黑龙江中医药大学,哈尔滨150040 [2]黑龙江中医药大学附属第二医院,哈尔滨150001 [3]黑龙江中医药大学附属第二医院哈南分院,哈尔滨150000 [4]黑龙江中医药大学附属第一医院,哈尔滨150040 [5]黑龙江中医药大学附属第一医院骨一科,哈尔滨150040

出　　处：《中华中医药杂志》2021年第12期7318-7323,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金面上项目(No.81874426);2020年度博士后科研启动金项目(No.LBH-Q20184);黑龙江省博士后科研基金面上项目(No.LBH-Z16194);黑龙江中医药大学校科研基金(No.2019BS10);黑龙江省大学生创新创业训练计划项目(No.201910228075)。

摘　　要：目的:探讨稳心丹对心肌缺血再灌注损伤(MIRI)大鼠心肌保护作用机制。方法:SPF级SD大鼠24只随机分为假手术组、模型组、稳心丹组,每组8只。采用方结结扎冠状动脉左前降支的方法建立MIRI模型,给予大鼠心肌缺血30min,再灌注60min。稳心丹组造模前按总生药量6g/kg灌胃14d;假手术组、模型组给予等体积的0.9%氯化钠溶液灌胃14d;生化仪检测CK-MB含量;TTC染色法计算心肌梗死面积;采用酶联免疫吸附法(ELISA)检测超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性、丙二醛(MDA)含量;流式细胞仪检测线粒体膜电位(Δψm)变化;采用原位末端转移酶标记(TUNEL)法测定细胞凋亡指数;透射电镜观察线粒体结构变化;蛋白免疫印迹法(Western Blot)测定细胞色素c(Cyt-c)蛋白含量;免疫组化法测定Cyt-c IOD值。结果:与假手术组比较,模型组血清中CK-MB含量升高、心肌梗死面积增大、MDA含量显著升高,SOD、NOS活性显著下降、线粒体膜电位显著下降、心肌细胞凋亡指数显著增高、Cyt-c的IOD值显著升高(P<0.01),Cyt-c的蛋白表达显著增加(P<0.05),心肌细胞线粒体出现肿胀,局部膜崩解、消失,部分出现内嵴溶解;与模型组比较,稳心丹组血清中CK-MB含量显著降低、心肌梗死面积显著降低、SOD、NOS活性显著升高(P<0.01),MDA含量显著降低(P<0.01)、心肌细胞凋亡指数显著降低、Cyt-c的IOD值显著下降(P<0.01),Cyt-c的蛋白表达显著降低(P<0.05),线粒体膜电位下降,心肌线粒体肿胀,内嵴部清晰,溶解现象减轻。结论:稳心丹可以通过抑制氧化应激反应,稳定心肌细胞线粒体膜电位,减少线粒体损伤,抑制心肌细胞凋亡发挥心肌保护作用。Objective: To explore the protective effect and mechanism of Wenxin Dan on myocardial ischemiareperfusion injury(MIRI) in rats. Methods: Twenty-four SPF SD rats were randomly divided into sham operation group, model group and Wenxin Dan group, 8 rats in each group. The MIRI model was established by ligating the left anterior descending coronary artery with square ligation. The rats were given 30 minutes of myocardial ischemia and 60 minutes of reperfusion.The Wenxin Dan group was given 6 g/kg for 14 days before modeling;the sham operation group and the model group were given intragastric saline for 14 days;biochemical analyzer to detect CK-MB content;calculate the area of myocardial infarction by TTC staining method;Enzyme-linked immunosorbent assay(ELISA) was used to detect superoxide dismutase(SOD) activity,nitric oxide synthase(NOS) activity, and malondialdehyde(MDA) content;Flow cytometry to detect changes in mitochondrial membrane potential(Δψm);TUNEL method for determining apoptosis index;Transmission electron microscope observation of mitochondrial structure changes;Western Blot was used to determine the protein content of cytochrome C;immunohistochemistry was used to determine the Cyt-C IOD value. Results: Compared with the sham operation group, the model group had increased serum CK-MB activity, increased myocardial infarction area, significantly increased MDA and ROS content, significantly decreased SOD activity, significantly decreased mitochondrial membrane potential, significantly increased myocardial cell apoptosis, the IOD value of Cyt-c is significantly increased(P<0.01), the protein expression of Cyt-c is significantly increased(P<0.05), the mitochondria of myocardial cells swell, the local membrane disintegrates and disappears, and part of the internal crest dissolves;Compared with the model group, the serum CK-MB activity in the Wenxin Dan group was significantly reduced(P<0.01),the myocardial infarction area was significantly reduced(P<0.01), the SOD activity was significantly increased, the

关 键 词：稳心丹 心肌缺血再灌注损伤 氧化应激 线粒体 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]