黄芪多糖对急性心肌梗死大鼠心室重构及miRNA-21的影响  

Effects of Astragalus polysaccharides on ventricular remodeling and miRNA-21 in rats with acute myocardial infarction

作　　者：董扬[1] 张芬[1] 李幸幸 吴杰 徐忠诚 DONG Yang;ZHANG Fen;LI Xing-xing;WU Jie;XU Zhong-cheng(Department of Cardiology,Jinhua People's Hospital,Jinhua 321000,China)

机构地区：[1]金华市人民医院心内二科,浙江金华321000

出　　处：《海南医学院学报》2021年第8期572-578,共7页Journal of Hainan Medical University

基　　金：浙江省基础公益研究计划项目(LGD19H020001)。

摘　　要：目的:探讨黄芪多糖(astragalus polysaccharides,APS)对心肌梗死后心肌重构及miR-21表达水平的影响。方法:选用specific pathogen free级雄性大鼠60只,随机分成假手术组、模型组及黄芪多糖低、中、高剂量组和阿托伐他汀组,每组10只。利用结扎冠状动脉左前降支的方法制备大鼠心肌梗死模型,造模后给予相应的药物干预4周。取各组大鼠心肌组织,分别采用苏木素-伊红(hematoxylin-eosin,HE)染色及Masson染色法观察心肌组织形态及胶原的变化,采用(enzyme linked immunosorbent assay,ELISA)法检测白细胞介素1β(interleukin-1β,IL-1β)、白细胞介素6(interleukin-6,IL-6)、白细胞介素-10(interleukin-10,IL-10)和肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的水平,采用逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction, RT-PCR)检测miR-21及基质金属蛋白酶-2 (matrix metalloproteinase-2,MMP2)、金属蛋白酶组织抑制剂-2(tissue inhibitor of matrix metalloproteinases-2,TIMP-2)、Ⅰ型胶原(collagen Ⅰ,Col-Ⅰ)、Ⅲ型胶原(collagen Ⅲ,Col-Ⅲ)mRNA的表达,采用Western blot检测Toll样受体4(toll like receptor 4,TLR4)、核因子κB p65(nuclear factor kappa-B p65,NF-κB p65)和髓样分化因子-88(myeloid differentiation factor 88,MyD88)蛋白的表达水平。结果:与模型组相比,黄芪多糖可以不同程度改善心肌组织病理形态,增加心肌组织中IL-10的水平,减少胶原染色面积和IL-1β、IL-6和TNF-α含量,差异具有统计学意义(P<0.05)。同时,黄芪多糖能显著降低心肌组织中MMP2、Col-Ⅰ和Col-Ⅲ mRNA的表达水平及MMP2/TIMP-2的比值,增加TIMP-2 mRNA和miR-21的表达丰度(P<0.05)。不仅如此,黄芪多糖还能显著降低大鼠心肌组织中TLR4、p-NF-κB p65及MyD88蛋白表达水平,与模型组比较差异均具有统计学意义(P<0.05)。结论:黄芪多糖可通过上调大鼠心肌组织中miR-21的表达水平,抑制TLR4/MyD88/NF-κB信号通路的激活,对急性心肌梗�Objective:To investigate the effect of Astragalus polysaccharides(APS)on myocardial remodeling and expression of miR-21 after myocardial infarction. Methods:Sixty SPF grade healthy male rats were divided into the sham operation group,the model group,the APS groups treated with APS at low,medium and high doses and the atorvastatin group,with 10 rats in each group. The left anterior descending coronary artery(LAD)was ligated to establish myocardial infarction model in rats,and the corresponding drug intervention was given for 4 weeks. The changes of myocardial morphology and collagen were observed by HE and Masson staining. The levels of IL-1β,IL-6,TNF-α and IL-10 were detected by ELISA. The mRNA expressions of miR-21,MMP2,TIMP-2,Col-Ⅰ,and Col-Ⅲ were also determined by RT-PCR. Moreover,the protein expressions of TLR4,MyD88 and NF-κB p65 were detected by Western blotting. Results:Compared with the model group,APS could improve the pathological morphology of myocardial tissue,increase the level of IL-10 in myocardial tissue,and reduce the staining area of collagen and the contents of IL-1β,IL-6 and TNF-α(P<0.05). At the same time,APS decreased the expression of MMP2,Col-Ⅰ and Col-Ⅲ mRNA and the ratio of MMP2/TIMP-2,and increased the expression of TIMP-2 mRNA and miR-21 significantly(P<0.05). Furthermore,APS significantly reduced the expression of TLR4,p-NF-κB p65 and MyD88 protein in the myocardial tissue of rats with myocardial infarction(P<0.05). Conclusion:APS can inhibit the activation of TLR4/MyD88/NF-κB signaling pathway by upregulating the expression of miR-21,which plays a therapeutic role in ventricular remodeling after acute myocardial infarction.

关 键 词：黄芪多糖 心肌梗死 心室重构 MIRNA-21 TLR4/MyD88/NF-κB 

分 类 号：R285.5[医药卫生—中药学]