桃红芪术软肝煎基于TGF-β/Smad信号通路逆转上皮-间质转化抗肝纤维化作用

Taohong Qizhu Ruangan Jian reverses epithelialmesenchymal transition via transforming growth factor beta/Smad signaling pathway

目的:观察桃红芪术软肝煎从(transforming growth factor beta,TGF-β)/Smad信号通路逆转上皮-间充质细胞转化(e p i t h e l i a lmesenchymal transition,EMT)抗肝纤维化的作用.方法:取对数生长期的HepG2细胞进行试验,将细胞分为7组:空白组、TGF-β诱导EMT组、TGF-β+中药[低剂量组(p e a c h stilbene low dose group,THQL)、中剂量组(peach stilbene middle dose group,THQM)、高剂量组(peach stilbene high dose group,THQH)]、TGF-β+扶正祛瘀胶囊组(Fuzheng Huayu recipe,FZHY组)、TGF-β+秋水仙碱组(c o l c h i c i n e g r o u p,QSXJ组),采用免疫荧光和Western blot方法检测E-cadherin、Vimentin、Smad2、TGF-βR1表达情况,并检测细胞上清液中的谷丙转氨酶(alanine aminotransferase,ALT)、谷草转氨酶(aspartate transaminase,AST)、甲胎蛋白(alpha fetoprotein,AFP)表达情况.结果:诱导组AFP诱导第3天浓度明显降低,差异具有统计学意义(P<0.05);用药干预后,用药组ALT、AST水平均低于模型组,差异具有统计学意义(P<0.05),THQL及TRQM的ALT水平均低于FZHY和QSXJ,差异具有统计学意义(P<0.05),THQM及THQH的AST水平均低于FZHY和QSXJ,差异具有统计学意义(P<0.05);桃红芪术软肝煎可以改善HepG2细胞被诱导发生EMT的形态,其中THQM组和THQH组形态改变较为明显;桃红芪术软肝煎可以上调E-cadherin表达,下调Smad2、TGF-βR1、Vimentin表达.结论:桃红芪术软肝煎可以通过作用于TGF-β/Smad信号通路逆转上皮-间质转化,从而起到抗肝纤维化的作用.

AIM： To explore whether Taohong Qizhu Ruangan Jian （TQRJ） reverses epithelial- mesenchymal transition （EMT） and the underlying mechanism. METHODS： HepG2 cells were divided into seven groups： a blank group, an EMT group, low-, medium- and high-dose TQRJ groups, a Fuzheng Huayu recipe group （FZHY group）, and a colchicine group （QSXJ group）. Irnmunofluorescence and Western blot were used to detect the expression of E-cadherin, Vimentin, Smad2, and transforming growth factor beta （TGF-β） R1. The levels of alanine aminotransferase （ALT）, aspartate transaminase （AST）, and alpha fetoprotein （AFP） in cell supernatants were also determined. RESULTS： AFP concentration significantly decreased after 3 d in the EMT group （P 〈 0.05）. After drug intervention, ALT and AST levels decreased significantly compared with the EMT group （P 〈 0.05）. ALT levels were significantly lower in the low- and medium- dose TQRJ groups than in the FZHY and QSXJ groups （P 〈 0.05）, and AST levels were significantly lower in the medium- and high- dose TQRJ groups than in the FZHY and QSXJ groups （P 〈 0.05）. TQRJ could improve EMT,especially in the medium- and high-dose TQRJ groups. TQRJ could increase E-cadherin expression and reduce the expression of Smad2, TGF-β R1 and Vimentin. CONCLUSION： TQRJ can reverse EMT possibly via the TGF-β/Smad signaling pathway and thus may has anti-liver fibrofic effects.