泡球蚴囊液对大鼠肝细胞TGF-βⅡ型受体及Smads信号蛋白表达的影响

Effect of Echinococcus multilocularis fluid on the expression of TGF-β receptorⅡ and Smads proteins inrat hepatocytes in vitro

目的观察泡球蚴(Em)囊液对Wistar大鼠肝细胞TGF-βⅡ型受体(TβRⅡ)及信号蛋白Smad2/3、Smad4及Smad7表达的影响。方法以Wistar大鼠作肝细胞供者,采用胶原酶肝脏原位灌流消化法分离,1×106接种于Ⅰ型胶原铺底35 mm培养皿,无血清培养20 h后,以泡球蚴囊液置换培养液培养2 h,提取蛋白,Western blot检测TGF-βⅡ型受体(TβRⅡ)、p-Smad2/3、Smad4及Smad7在肝细胞的表达。结果 Western blot显示,泡球蚴囊液处理2 h后,大鼠肝细胞内TβRⅡ、p-Smad2/3、Smad4及Smad7分别为4.71±0.73、2.37±0.34、2.08±0.23和0.19±0.05,对照组分别为1.55±0.21、0.42±0.09、0.37±0.07和0.59±0.12,差异均有统计学意义(P<0.05或P<0.01)。结论泡球蚴囊液对大鼠肝细胞TβRⅡ、p-Smad2/3、Smad4蛋白表达均有上调作用,而对Smad7蛋白表达具有下调作用,提示泡球蚴侵染宿主过程中通过影响TGF-/βSmads信号通路而造成宿主肝损伤。

Objective To observe the expression of TGF-β receptor Ⅱ（TβRⅡ） and Smad signaling proteins in rat hepatocytes treated with Echinococcus multilocularis fluid（EmF）.Methods Hepatocytes were isolated from Wistar rats by collagenase digestion and were cultured in 35-mm culture dishes coated with collagen Ⅰ at 1×10^6.After culturing in a serum-free conditioned medium for 20 hours,hepatocytes were incubated with EmF for 2 hours.The expression of TβRⅡ,pSmad2/3,Smad4,and Smad7 was analyzed by Western blotting.Results Two hours after treatment with EmF,the expression of TβRⅡ,p-Smad2/3,Smad4,and Smad7 was 4.71±0.73,2.37±0.34,2.08±0.23,and 0.19±0.05 in the EmF group and 1.55±0.21,0.42±0.09,0.37±0.07 and 0.59±0.12（P〈0.05 or P〈0.01） in the control group.Conclusion The expression of TβRⅡ,p-Smad2/3,and Smad4 protein significantly increased,with an abnormal decrease in Smad7 in the rat hepatocytes treated with EmF.Disruption of the TGF-β/Smad signaling pathways may play an important role in the invasion and spread of Em.