黄芪多糖通过Toll样受体4/核转录因子-κB信号通路抑制异丙肾上腺素诱导的心肌细胞肥大  

Astragalus polysaccharides inhibites cardiomyocyte hypertrophy induced by isoproterenol via Toll-like receptor 4/nuclear factor-κB signal pathway

作　　者：张静[1] 王洪新[1] 杨娟[1] 鲁美丽[1] 李胜陶[1] 喻晓春[2] 

机构地区：[1]辽宁医学院药物研究所心血管药物研究重点实验室,辽宁锦州121001 [2]中国中医科学院实验中心

出　　处：《中华高血压杂志》2014年第2期157-162,共6页Chinese Journal of Hypertension

基　　金：国家自然科学基金(30973898/C190702);辽宁省自然科学资金项目(201102141)

摘　　要：目的探讨黄芪多糖抑制异丙肾上腺素(Iso)诱导心肌细胞肥大的作用及机制。方法原代培养SD乳鼠心肌细胞,用Iso 10μmol/L诱导心肌细胞肥大,观察核转录因子-κB(NF-κB)特异性抑制剂BAY11-7082及不同浓度黄芪多糖对肥大心肌细胞的影响。用消化分离法及计算机图像分析系统测细胞体积;考马斯亮蓝法测心肌细胞总蛋白含量;酶联免疫吸附试验(ELISA)检测细胞外液中肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的含量;逆转录聚合酶链反应(RT-PCR)法检测心房钠尿肽(ANP)mRNA和Toll样受体4(TLR4)mRNA的表达;蛋白免疫印迹(Western blot)法检测心肌细胞P65、NF-κB抑制蛋白IκBα和TLR4蛋白的表达。结果与正常对照组相比,Iso组心肌细胞体积增大88.3%,总蛋白含量增加55.3%,ANP mRNA表达增高,炎症因子TNF-α和IL-6的含量分别增加200%和65.9%,TLR4mRNA表达增高,P65和TLR4蛋白含量增多及IκBα蛋白含量减少。黄芪多糖和BAY11-7082均能有效抑制Iso诱导的心肌细胞肥大,表现为体积减小,总蛋白含量降低,ANP mRNA表达降低;均能有效减少炎症反应,表现为细胞外液中TNF-α和IL-6明显减少;均使TLR4mRNA表达降低,P65和TLR4蛋白含量减少及IκBα蛋白含量增多,且黄芪多糖呈一定的剂量依赖性。结论黄芪多糖对Iso诱导的乳鼠心肌细胞肥大有保护作用,其机制可能与抑制TLR4/NF-κB炎症信号通路有关。Objective To discuss the effects and mechanism of astragalus polysaccharides（APS） on cardiomyocyte hypertrophy induced by isoproterenol {Iso）. Methods Cardiac myocytes from SIS） neonatal rat were primarily cul- tured and induced hypertrophy by Iso （10 μmol/L）. The effects of NF-kB＇s inhibitor BAYll-7082 and different concentrations of APS on hypertrophic cardiomyocyte were observed. The volume of cardiomyocytes was measured by digestive isolation and computer photograph analysis system, the total protein content by coomassie brilliant blue （Bradford}, the expression of tumor necrosis factor a （TNF-~} and interleukin-6 （IL-6} by enzyme-linked immu- nosorbent assay （ELISA）, the expression of atrial natriuretic peptide（ANP） mRNA and Toll-like receptor 4（TLR4） mRNA by inverse transcription polymerase chain reaction （RT-PCR）, and the expression of myocardial cell P65, I^Ba and TLR4 protein expression by Western blot. Results Compared with the normal control group, the cardio- myocyte size, total protein content,content of TNF-a and IL-6 in Iso group increased by 88.3%, 55.3%, 200~, 65.9~, respectively, along with the expression of ANP mRNA, TLR4 mRNA, P65 protein and TLR4 protein, while the protein expression of I^Ba decreased. BAY11-7082 and APS could inhibit the cardiomyocyte hypertrophy induced by Iso, and decrease the inflammatory response and the expression of TLR4 mRNA, with that manifestation the cell volume, total protein content, expression of ANP mRNA, TNF-a and IL-6 in extracellular fluid, P65 and TLR4 protein decreased, and IkBa protein increased. APS had a dose-dependent inhibitory effect. Conclusion APS could inhibit cardiomyocyte hypertrophy induced by Iso, with the mechanism that may relate to TLR4/NF-kB in- flammatory signaling pathway.

关 键 词：黄芪多糖 异丙肾上腺素 心肌细胞 TOLL样受体4 核转录因子-KB 炎症 

分 类 号：R285.5[医药卫生—中药学]