丹参通络解毒汤对心肌缺血再灌注损伤大鼠的保护作用及自噬和血管生成因子的影响  

Protective effects of DanShenTongLuo-detoxification decoction on rats with myocardial ischemia-reperfusion injury and the effects of autophagy and angiogenic factors

作　　者：李彩云 刘仕成 李鑫辉 LI Cai-yun;LIU Shi-cheng;LI Xin-hui(College of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;Second Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410005,China)

机构地区：[1]湖南中医药大学中医学院,湖南长沙410208 [2]湖南中医药大学第二附属医院,湖南长沙410005

出　　处：《时珍国医国药》2024年第2期293-296,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金面上项目(82074392);湖南省中医药管理局重点项目(A2023013);湖南省自然科学基金联合基金项目(2023JJ50036)。

摘　　要：目的探讨丹参通络解毒汤(DSTL)对大鼠心肌缺血再灌注损伤(MIRI)的保护作用及自噬和血管生成因子的影响。方法将雄性SD大鼠随机平均分成5组:假手术组(Sham组)、MIRI模型组(MIRI组)、MIRI模型+DSTL组(DSTL组)、MIRI模型+自噬抑制剂3-甲基腺嘌呤组(3-MA组)以及MIRI模型+自噬激动剂雷帕霉素组(Rap组),每组10只。采用苏木精-伊红(HE)染色评估心肌的病理变化,酶联免疫吸附试验(ELISA)检测心肌细胞损伤标志物肌酸激酶MB同功酶(CK-MB)和心肌肌钙蛋白I(CTnI)的水平,免疫组织化学染色检测Beclin1的表达和分布,蛋白免疫印迹(WB)测定p62、内皮一氧化氮合成酶(eNOS)和血管内皮生长因子(VEGF)的表达水平。结果MIRI组大鼠在显微镜下显示严重的心肌细胞坏死和炎症浸润,CK-MB、CTnI和Beclin1的水平明显升高(P<0.01),p62、eNOS和VEGF的水平显著降低(P<0.01),而DSTL组的大鼠显示较轻微的病理变化,CK-MB、CTnI和Beclin1的水平下降(P<0.01),p62、eNOS和VEGF的水平升高(P<0.01);DSTL组的Beclin1和p62的水平介于3-MA组和Rap组(P<0.01),而eNOS和VEGF的水平最高(P<0.01)。结论DSTL能适度抑制自噬,促进血管生成因子的产生,从而减轻MIRI,保护大鼠心肌细胞。Objective To investigate the protective effects of DanShenTongLuo-detoxification decoction(DSTL)on rats with myocardial ischemia-reperfusion injury(MIRI)and the effects of autophagy and angiogenic factors.Methods Male Sprague-Dawley(SD)rats were randomly and evenly assigned into five groups:sham-operated group(Sham group),MIRI model group(MIRI group),MIRI model+DSTL group(DSTL group),MIRI model+autophagy inhibitor 3-methyladenine group(3-MA group),and MIRI model+autophagy agonist rapamycin group(Rap group),with ten rats in each group.Hematoxylin-eosin(HE)staining was performed to evaluate the pathological changes in cardiac muscle.Enzyme-linked immunosorbent assay(ELISA)was carried out to detect the levels of markers of cellular damage,including creatine kinase MB isoenzyme(CK-MB)and cardiac troponin I(CTnI).In addition,immunohistochemical staining was performed to analyse the expression and distribution of Beclin1.Western blotting(WB)was conducted to measure the expression levels of p62,endothelial nitric oxide synthase(eNOS)and vascular endothelial growth factor(VEGF).Results Rats in the MIRI group showed severe myocardial cell necrosis and inflammatory infiltration under the microscope,with significantly increased levels of CK-MB,CTnI and Beclin1(P<0.01)and significantly decreased levels of p62,eNOS and VEGF(P<0.01),whereas rats in the DSTL group showed milder pathological changes,with increased levels of CK-MB,CTnI and Beclin1 levels were decreased(P<0.01)and p62,eNOS and VEGF levels were increased(P<0.01).Levels of Beclin1 and p62 in the DSTL group were intermediate between the 3-MA group and the Rap group(P<0.01),while eNOS and VEGF levels were highest(P<0.01).Conclusion DSTL moderately inhibits autophagy and promotes angiogenic factor production,thereby attenuating MIRI and protecting cardiomyocytes in rats.

关 键 词：丹参通络解毒汤 心肌缺血再灌注损伤 自噬 血管新生因子 

分 类 号：R285.5[医药卫生—中药学]