氧化苦参碱抑制DBTC刺激的胰腺腺泡细胞中上皮间质转化相关蛋白及TβRⅡ和p-Smad2/3表达

Oxymatrine inhibits epithelial-mesenchymal transition and the related protein expressions of TβRⅡ and p-Smad2/3 induced by Dibutyltin dichloride in pancreatic acinar cells

目的探讨二氯二丁基酯(Dibutyltin dichloride,DBTC)及氧化苦参碱(oxymatrine,OM)对胰腺腺泡细胞中上皮间质转化(epithelial-mesenchymal transition,EMT)相关蛋白和TGFβ1/Smad信号通路影响。方法体外常规传代培养AR42J细胞。CCK-8法检测0、0.1、1、10、100μmol/L的DBTC对AR42J细胞毒性作用。Western Blot法检测0、0.1、1、10μmol/L的DBTC刺激AR42J细胞72h后EMT相关蛋白E-cadherin、Vimentin及Snail1表达。Western Blot法检测对照组(Control组)、模型组(DBTC:10μmol/L)、OM治疗组(DBTC:10μmol/L;OM:1mg/m L)及药物对照组(OM:1 mg/m L)中E-cadherin、Vimentin、Snail1及转化生长因子β1Ⅱ型受体(TβRⅡ)、p-Smad2/3的表达。结果浓度低于10μmol/L的DBTC对AR42J细胞无明显的毒性作用。1μmol/L及10μmol/L DBTC刺激AR42J细胞可下调E-cadherin同时上调Vimentin、Snail1的表达。与对照组相比,10μmol/L DBTC刺激AR42J细胞72 h后还可上调TβRⅡ、p-Smad2/3表达,OM可逆转上述蛋白的表达。结论 DBTC可诱导胰腺腺泡细胞发生EMT,OM对此有抑制作用,其机制可能与TGFβ1/Smad通路有关。

Objective To explore the effects of Dibutyltin dichloride （DBTC） and oxymatrine （OM） on epithe- lial- mesenchymal transition （EMT） and the related protein expressions of TGFβ1/Smad signaling pathway in pancreatic acinar cells. Methods AR42J cells were cultured in vitro and treated with DBTC for 72 hours at different concentrations. The cytotoxicity of DBTC on AR42J cells was measured by CCK - 8 assay. The protein expres- sions of E - cadherin, Vimentin, Snaill ,T[3R ] and p - Smad2/3 treated by DBTC with or without OM in AR42J cells were detected by Western Blotting. Results DBTC had no significant cytotoxicity on AR42J cells at less than 10 μmol/L concentration. DBTC （ 1 p, mol/L or 10 μmol/L） could downregulate E - cadherin and upregulate Vi- mentin and Snaill in AR42J cells. Compared with control group, DBTC （10μmol/L） stimulated AR42J cells for 72 hours and also upregulated TI3R I1 and p -Smad2/3, whereas OM reversed the alterations. Conclusion DBTC induces EMT, which could be inhibited by OM, via TGFβ/Smad signaling pathway in pancreatic acinar cells.