氧化苦参碱下调p38MAPK磷酸化及改善胶原沉积抑制TGF-β1诱导的CFBs增殖  

OMT inhibited TGF-β1-induced cardiac fibroblast proliferation via down-regulating p38MAPK phosphorylation in vitro

作　　者：肖海[1,2] 徐旖旎[1] 罗红[1] 陈妍[1,2] 张彦燕[1] 陶玲[1] 江滟[1] 沈祥春[1,2] 

机构地区：[1]贵州医科大学天然药物资源优效利用重点实验室,贵州贵阳550004 [2]贵州医科大学中药药理教研室,贵州贵阳550004

出　　处：《中国中药杂志》2015年第11期2168-2173,共6页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(81173586);教育部新世纪优秀人才支持计划(NCET-13-0747);贵州省省长资金(黔科教办[2011]28号);贵州省高层次创新型人才(黔科合人才[2015]4029号)

摘　　要：目的:研究氧化苦参碱(oxymatrine,OMT)对TGF-β1诱导心肌成纤维细胞(CFBs)增殖的作用,并分析可能的作用机制。方法:试验分为空白对照组(无血清DMEM)、模型组(20μg·L-1TGF-β1)、OMT低剂量组(OMT-L组,1.89×10-4mol·L-1)、OMT中剂量组(OMT-M组,3.78×10-4mol·L-1)、OMT高剂量组(OMT-H组,7.56×10-4mol·L-1)和SB203580(p38MAPK阻断剂,1×10-5mol·L-1)。免疫细胞化学法鉴定CFBs中波形蛋白,测定TGF-β1诱导CFBs 24 h后α-SMA的表达情况;采用MTT法分析CFBs增殖的抑制作用;苦味酸天狼星红染色观察Ⅰ型胶原和Ⅲ型胶原的沉积;Western blot分析p38MAPK,p-p38MAPK,Ⅰ型胶原和Ⅲ型胶原蛋白的表达。结果:MTT结果提示3.78×10-4,7.56×10-4mol·L-1的OMT对TGF-β1诱导的CFBs异常增殖具有抑制作用(P<0.01);α-SMA免疫细胞化学实验提示OMT能够抑制CFBs-my FBs转换;苦味酸天狼星红染色显示OMT能明显减少Ⅰ型胶原和Ⅲ型胶原的沉积;Western blot表明OMT可以显著抑制TGF-β1诱导Ⅰ型胶原和Ⅲ型胶原的沉积及p38MAPK的磷酸化(P<0.01)。结论:OMT可抑制TGF-β1诱导CFBs增殖的作用并抑制相关胶原蛋白的表达,改善心肌纤维化,其机制可能与抑制p38MAPK磷酸化有关。Objective： To investigate the inhibitory effects of OMT on TGF-β1-induced CFBs proliferation, and then explore the mechanism. Method： The experiment was randomly divided into 6 groups as following： control group （serum free DMEM）, model group （20 μg·L^-1 TGF-β1）, OMT low dose group （1.89×10^-4 mol·L^-1＋20 μg·L-1TGF-β1）, OMT medium dose group （3.78×10^-4 mol·L^-1＋20 μg·L^-1TGF-β1）,OMT high dose group（7.56×10-4 mol·L^-1＋20 μg·L^-1TGF-β1）,SB203580 group （p38MAPK blocking agent,1×10^-5 mol·L^-1 ＋20 μg·L^-1TGF-β1）.Vimentin of CFBs was identified by immunocytochemical methods, α-SMA of myFBs as well. Inhibitory effects of OMT on CFBs proliferation was detected by the MTT assay. Picric acid Sirius red staining was analyzed collagen type Ⅰ and collagen type Ⅲ deposition. Western blot was determined the expression of p38MAPK, p-p38MAPK, collagen type Ⅰ and collagen type Ⅲ. Result： MTT results showed that OMT significantly inhibited CFBs proliferation induced by TGF-β1（P〈0.01）;α-SMA immunocytochemical experiments suggested that OMT could protect against the CFBs proliferation. OMT could significantly decrease the deposition of collagen type Ⅰ and collagen type Ⅲ by Western bloting and picric acid Sirius red staining. Western blot results showed that TGF-β1 enhanced p38MAPK phosphorylation, however OMT attenuated the phosphorylation of p38MAPK induced by TGF-β1（P〈0.01）. Conclusion： OMT can inhibit the CFBs proliferation induced by TGF-β1, and its mechanism may be involved in inhibiting p38MAPK phosphorylation.

关 键 词：氧化苦参碱 TGF-Β1 心肌纤维化 心肌成纤维细胞 胶原蛋白 P38MAPK 

分 类 号：R285[医药卫生—中药学]