抗纤软肝颗粒含药血清对肝卵圆细胞系WB-F344上皮间质转化的影响

Effect of Serum Containing Kangxian Ruangan Granule on Epithelial-mesenchymal Transformation of Hepatic Oval Cell Line WB-F344

目的研究抗纤软肝颗粒含药血清(KXRG)对N-甲基-N-硝基-N-亚硝基胍(MNNG)诱导肝卵圆细胞系WB-F344上皮间质转化的影响。方法采用MNNG干预WB-F344细胞系制备上皮间质转化(EMT)细胞模型,实验组用高、中、低不同浓度抗纤软肝颗粒含药血清(KXRG)刺激细胞,正常组、模型组加等量大鼠空白血清;采用CCK-8法检测细胞的增殖能力,Western blot法检测E-cadherin,N-cadherin,Vimentin蛋白的表达;酶联免疫吸附实验(ELISA)检测细胞上清中甲胎蛋白(AFP)含量。结果与正常组比较,模型组细胞OD值、细胞上清中AFP浓度、N-cadherin、Vimentin蛋白表达水平均明显升高(P<0.05);而E-cadherin蛋白表达水平降低(P<0.05)。与模型组相比,含药血清各组细胞OD值、细胞上清中AFP浓度、N-cadherin、Vimentin蛋白表达水平均显著下降(P<0.05),且含药血清浓度越高,上述指标下降越明显;而E-cadherin蛋白表达水平明显上升(P<0.05),且含药血清浓度越高,Ecadherin蛋白表达水平上升越明显。结论抗纤软肝颗粒含药血清可以抑制大鼠肝卵圆细胞WB-F344细胞的上皮间质转化。

Objective To study the effect of serum containing Kangxian Ruangan granule(KXRG)on n-methyl-n-nitro-n-nitrosoguanidine(MNNG)-induced epithelial-mesenchymal transformation of hepatic oval cell line WB-F344.Methods WB-F344 cell line was treated with MNNG.The experimental group was treated with KXRG of high,medium and low concentrations,and the normal group and model group were treated with the same amount of blank serum.The expression of E-cadherin,N-cadherin and Vimentin was detected by Western blot,and AFP was detected by enzyme-linked immunosorbent assay(Elisa).Results Compared with the normal group,the OD value,AFP concentration,N-cadherin and Vimentin expression in model group were significantly increased(P<0.05),while E-cadherin protein expression was significantly decreased(P<0.05).Compared with model group,OD value,AFP concentration,N-cadherin and Vimentin in drug-containing serum group were significantly decreased(P<0.05),and the higher the concentration of drug-containing serum was,the more obvious the decrease was.The expression of E-cadherin increased significantly(P<0.05),and the higher the concentration of drugcontaining serum was,the higher the expression of E-cadherin was.Conclusion KXRG can inhibit the epithelial-mesenchymal transformation of WB-F344 cells.