苦参碱诱导大鼠肝卵圆细胞株WB-F 344细胞分化中Wnt-1信号通路的作用

Role of Wnt-1 signal pathway in matrine-induced differentiation of rat hepatic oval cell line WB-F 344

目的:研究苦参碱干预体外培养的大鼠肝卵圆细胞分化中Wnt-1信号通路的作用.方法:体外培养大鼠肝卵圆细胞,分别用不同浓度的苦参碱(0.001、0.01、0.05、0.2、0.5、1、1.5、2g/L)处理卵圆细胞,HE染色观察细胞的形态变化,免疫细胞化学检测细胞内甲胎蛋白、白蛋白、Wnt-1信号蛋白的表达.结果:苦参碱对肝卵圆细胞的生长有抑制作用,且苦参碱浓度越大,作用时间越长,抑制作用越明显,0.05、0.2、1g/L的苦参碱作用72h后,对卵圆细胞的抑制率分别为24.16%±2.03%、40.25%±3.92%和67.31%±6.04%.0.01g/L的苦参碱即可使肝卵圆细胞的形态发生变化,细胞核变大、变圆,核浆比例减小,双核细胞增多;免疫细胞化学结果显示,0.2g/L的苦参碱能够使肝卵圆细胞的ALB表达增高,AFP的表达降低,Wnt-1信号蛋白的表达量降低.结论:苦参碱可以抑制Wnt信号通路的传导使卵圆细胞向着成熟肝细胞的趋势分化.

AIM：To study the role of Wnt-1 signal pathway in matrine–induced differentiation of rat oval cell line WB-F 344. METHODS：Hepatic oval cells were treated with different concentrations of matrine（0.001,0.01,0.05, 0.2,0.5,1,1.5,2 g/L）.The morphological changes were measured using HE staining and protein expressions of the albumin,AFP and Wnt-1 were detected using immunohistochemistry. RESULTS：Seventy two hours after the treat- ment of matrine at concentrations of 0.05,0.2 and 1 g/L,the inhibitory rates were 24.16% ±2.03%,40.25%±3.92%and 67.31%±6.04%,respectively.0.01 g/L matrine induced morphological changes of hepatic oval cell,including diminished karyoplasmic ratio,bigger and rounder karyon and increased dikaryocyte.Im- munohistochemical results showed that the expression of albumin was significantly increased, the expressions of AFP and Wnt-1 protein were significantly decreased compared with normal hepatic oval cell after treatment of 0.2 g/L matrine. CONCLUSION：Matrine induces rat hepatic oval cells to differentiate into mature hepatocytes, and inhibits the conduction of Wnt signal pathway.