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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。
Objective: To investigate the effect of bone morphogenetic protein-4 (BMP-4) on proliferation and differentiation of rat hepatic stem cell line WB-F344. METHODS: WB-F344 cells were treated with 50 ng / mL recombinant BMP-4 and BMP-4 antagonist Noggin (200 ng / mL) blocked the effect of BMP-4. The effect of BMP-4 on the proliferation of WB-F344 was detected by MTT colorimetric assay at different time points. The ultrastructure of cells was observed by transmission electron microscope. WB-F344 cells may differentiate phenotype-specific molecular marker mRNA expression. Results: The results of cell proliferation assay showed that the cell absorbance of BMP-4 intervention group was higher than that of Noggin treatment group and blank control group at different time points. WB-F344 cells, which were observed under transmission electron microscope after BMP-4 intervention, had well-differentiated ultrastructural features of epithelial cells. RT-PCR showed significant mRNA expression of hepatocyte-specific molecular markers of hepatic stem cells after BMP-4 induced WB-F344 differentiation. Conclusion: BMP-4 can promote the proliferation of hepatic stem cells WB-F344 and induce its directional differentiation into hepatocytes.