Toll样受体4/NF-κB信号通路参与黄芪多糖对肿瘤坏死因子α诱导的乳大鼠心肌细胞肥大的抑制作用  

Astragaluspolysaccharides ameliorate hypertrophy of cardiomyocytes induced by tumor necrosis factor-α via TLR4/NF-κB signaling pathway

作　　者：梁灵君[1] 王洪新[1] 孙雪芳[1] 喻晓春[2] 鲁美丽[1] 顾洁莹[1] 何海洋[1] 

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,北京100700

出　　处：《中国药理学与毒理学杂志》2013年第2期168-173,共6页Chinese Journal of Pharmacology and Toxicology

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(APS)对肿瘤坏死因子α(TNF-α)诱导的乳大鼠心肌细胞肥大的保护作用及机制。方法 原代培养乳大鼠心肌细胞分别加入APS 25,50和100 mg.L-1作用30 min后再加入TNF-α50μg.L-1作用48 h。考马斯亮蓝法测定心肌细胞蛋白含量;消化分离法及计算机图像分析系统检测细胞体积;RT-PCR检测心肌心钠素(ANP)和Toll样受体4(TLR4)的mRNA表达;Western印迹法检测心肌细胞p65和IκBα的蛋白表达;ELISA法检测细胞外液白细胞介素1β(IL-1β)的含量。结果 与正常对照组相比,TNF-α组心肌细胞蛋白含量增加了52.8%,细胞体积增大了66.7%,ANP和TLR4 mRNA分别增加了60.3%和68.7%,p65蛋白表达增加了50.5%,IκBα蛋白表达减少了43.1%,IL-1β表达增加了59.0%,差异均有统计学意义(P<0.01)。与TNF-α模型组相比,APS 50,100 mg.L-1和IκBα阻断BAY11-7082 5μmo.l L-1组蛋白含量、细胞体积、ANP mRNA、TLR4 mRNA、IL-1β和p65蛋白表达显著降低(P<0.05),IκBα蛋白表达明显增高(P<0.05);APS 25 mg.L-1组蛋白含量、细胞体积和ANP mRNA表达显著性降低(P<0.05)。结论 APS对TNF-α诱导的乳大鼠心肌细胞肥大有保护作用,其机制可能与抑制TLR4/NF-κB信号通路有关。OBJECTIVE To observe the inhibitory effect and signal transduction of Astragalus poly- saccharide （APS） on hypertrophic myocardial cells induced by tumor necrosis factor-α （TNF-α） in neo- natal rats. METHODS The cardiac myocytes were treated with APS 25, 50 and 100 mg·L-1 for 30 min, before TNF-a 50 pg · L-1 was added and incubated for 48 h. The total protein content was assayed by the method of Bradford. The cardiomyocyte volume was measured by computer photograph analysis system. The expression of atrial natriuretic peptide （ANP） and Toll-like receptor 4 （TLR4） mRNA were determined by RT-PCR. The expression of p65 and inhibitor-KBα（IKBα） was measured by Western blotting, and the expression of interleukin-lβ （IL-Iβ） was determined by ELISA. RESULTS Compared with normal control group, the total protein content, cell size, ANP mRNA, TLR4 mRNA, p65 and IL-1β were increased by 52.8%, 66.7%, 60.3%, 68.7%, 50.5%, and 59.0%, respectively, and the expression of IKBa was decreased by 43.1% in TNF-α group. Compared with TNF-a group, APS 50, 100 mg· L-1 and BAY11-7082 5 pmol. L-1 remarkably down-regulate the overexpression of the total protein content, cell size, ANP mRNA, TLR4 mRNA, p65 and IL-Iβ（ P 〈0.05）, but increased the ex- pression of IKBa （P 〈0.05）. APS 25 mg·L-1 could decrease protein content, cell size and ANP mRNA expression （P〈0.05）. CONCLUSION APS has a protective effect on TNF-a-induced cardiac hyper trophy, partially by attenuating the inflammatory effect and high expression of TLR4/NF-KB.

关 键 词：黄芪多糖 肿瘤坏死因子Α 心肌肥大 炎症反应 Toll样受体4/NF-κB信号通路 

分 类 号：R285.5[医药卫生—中药学]