Objective To determine the regulation effect of bone morphogenetic protein-4 (BMP-4) on the proliferation and differentiation of rat hepatic precursor cells. Methods We used Noggin (200 ng/mL) as the function blocking control of BMP-4, and the hepatic precursor cells of WB-F344 were treated with recombinant BMP-4 at 50 ng/mL at different time points. The proliferation of WB-F344 cells were tested by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The ultrastructural characters of differentiated WB-F344 cells regulated by BMP-4 were observed under a transmission electron microscope. RT-PCR was used to examine mRNA expression of specific molecular markers for different cellular phenotypes potentially differentiated from the WB-F344 cells. Results At different time points, the absorbance values in the BMP-4 treatment groups were higher than those in the control groups of Noggin and blank treatment ( P ＜ 0.01 ). The WB-F344 cells treated with BMP-4 exhibited typical ultrastructural characters of well-differentiated epithelial cells.The hepatocyte mRNA markers were more significantly promoted in the differentiated WB-F344 cells in the BMP-4 treatment group than those in the other 2 control groups. Conclusion BMP-4 can promote the proliferation and directional differentiation towards hepatocytes of rat hepatic precursor cells of WB-F344.%目的:探讨骨形态发生蛋白-4(BMP-4)对大鼠肝干细胞系WB-F344的增殖及分化的影响.方法:采用50 ng/mL的基因重组BMP-4作用于WB-F344细胞,以BMP-4拮抗剂Noggin(200 ng/mL)阻断BMP-4的作用为对照.在实验不同时间点,通过四甲基偶氮唑盐(MTT)比色法检测BMP-4对WB-F344增殖的影响,透射电镜下观察细胞的超微结构特征,RT-PCR检测各试验组中WB-F344细胞可能分化表型特异性分子标志物的mRNA表达.结果:细胞增殖实验显示:在不同作用时间点,BMP-4干预组的细胞吸光值均高于Noggin处理组和空白对照组.透射电镜下可观察到BMP-4干预后的WB-F344细胞具有分化良好的上皮细胞的超微结构特征.RT-PCR显示BMP-4诱导WB-F344分化后,肝干细胞的肝细胞特异性分子标志物的mRNA显著表达.结论:BMP-4可促进肝干细胞WB-F344的增殖,并诱导其向肝细胞定向分化.
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