苦参碱含药血清对大鼠肝干细胞的影响

The effect of matrine drug serum on rat liver stem cell in vitro

目的研究苦参碱含药血清对大鼠肝干细胞系WBF-344细胞的增殖及AFP、Notch1蛋白和mRNA表达的影响。方法用MTT法检测苦参碱含药血清对WBF-344细胞增殖的影响,免疫细胞化学及RT-PCR方法检测苦参碱含药血清对WBF-344细胞AFP、Notch1蛋白和mRNA表达的影响。结果5%、10%、20%浓度的苦参碱含药血清作用于WBF-344细胞24h、48h、72h均有不同程度的抑制增殖作用,且存在时间、剂量依赖性。正常培养WB-F344细胞有少量AFP、Notch1蛋白和mRNA表达,肝癌前病变模型组血清培养后AFP、Notch1蛋白和mRNA表达上调,苦参碱含药血清培养后AFP、Notch1蛋白和mRNA表达较模型组血清AFP、Notch1蛋白和mRNA表达下调。结论肝癌前病变模型组血清可能具有诱导WBF-344细胞向肝癌细胞方向分化的作用,苦参碱含药血清可逆转此作用,以上作用与调节Notch1表达关系密切。

Objective To explore matrine drug serum's effect on proliferation of rat liver stem cell and expression of AFP,Notchl protein and mRNA.Methods MTT method was used to examine the proliferation effect of matrine drug serum on rat liver stem cell.The immunocytochemistry stain method was used to study the expression of AFP,Notch1 protein and mRNA.Results The 5%,10%,20% matrine drug serum can inhibit proliferation of WBF-344 cells in 24 h,48 h,72 h,the inhibitory function depended on time and concentration.Expression of AFP,Notch1 protein and mRNA in control group were low and were up-regulated by hepatic precancerous lesion model sera.Expression in matrine drug serum group was lower than that in hepatic precancerous lesion model serum group.Conclusions The hepatic precancerous lesion model sera may induce WBF-344 into hepatoma carcinoma cell,matrine drug serum can reverse this effection,which was related to regulation of Notchl expression.