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目的检测肝细胞刺激因子肝细胞生成素Cn(hepatopoietin Cn,HPPCn)作用肝祖细胞的功能。方法利用MTT法、AnnexinⅤ和PI荧光染色法分别检测WB-F344细胞增殖及其抗凋亡能力,Transwell法检测细胞迁移能力。建立小鼠2AAF-部分肝切除(partial hepatectomy)模型,分析高表达HPPCn对肝干/祖细胞增殖和迁移的影响。结果重组HPPCn蛋白能促进WB-F344细胞的增殖及细胞迁移,低浓度HPPCn能抑制WB-F344细胞的凋亡,并激活WB-F344细胞中Sph K1、Erk1/2以及Stat3信号通路。动物模型检测结果显示,肝特异HPPCn转基因小鼠部分肝切除后,肝卵圆细胞较对照组明显增多,肝再生能力增强。结论 HPPCn能促进肝干/祖细胞增殖和存活,并通过促进肝卵圆细胞增生,提高肝再生能力。
Objective To investigate the function of hepatopoietin Cn (HPPCn) on hepatocyte progenitor cells. Methods The proliferation and anti-apoptotic ability of WB-F344 cells were detected by MTT, Annexin Ⅴ and PI staining respectively. The cell migration was assayed by Transwell assay. Mouse 2AAF-partial hepatectomy model was established to analyze the effect of high expression of HPPCn on the proliferation and migration of hepatic stem / progenitor cells. Results Recombinant HPPCn protein could promote the proliferation and cell migration of WB-F344 cells. Low concentrations of HPPCn could inhibit the apoptosis of WB-F344 cells and activate the Sph K1, Erk1 / 2 and Stat3 signal pathways in WB-F344 cells. Animal model test results showed that after partial hepatectomy in liver-specific HPPCn transgenic mice, hepatic oval cells were significantly increased compared with the control group, and liver regeneration ability was enhanced. Conclusion HPPCn can promote the proliferation and survival of hepatic stem / progenitor cells and improve hepatic regeneration by promoting hepatic oval cell proliferation.