粒细胞集落刺激因子保护急性损伤人胎肝细胞的实验研究

目的建立D-半乳糖胺(D-galactosamine,D-GalN)对人胎肝细胞急性损伤的模型,观察粒细胞集落刺激因子(Granulocyte-colony stimulating factor,G-CSF)对人胎肝细胞损伤的保护作用。方法分别用梯度浓度的D-GalN和不同的作用时间孵育人胎肝细胞,用四唑盐比色法(MTT法)检测细胞活性,以确定最佳的人胎肝细胞急性损伤造模条件。将胎肝细胞分为4组进行不同处理:第1组为空白对照组,第2组(G组)用G-CSF处理正常细胞,第3组(ND组)和第4组(GD组)都用D-GalN进行损伤造模,但GD组加入G-CSF作为治疗,第3组加入等量的0.9%氯化钠溶液作为实验对照。最后用MTT法和乳酸脱氢酶(LDH)释放量检测各组细胞活性。结果当D-GalN浓度为10 mg/ml,作用时间为12 h时,可以杀伤90%以上的人胎肝细胞,并且可以保证有足够的药物反应时间。空白对照组和G组的细胞活性差异无统计学差异,但GD组细胞活性明显高于ND组(P0.05)。结论 D-GalN对人胎肝细胞急性损伤的造模条件为D-GalN 10 mg/ml作用12 h。G-CSF对D-GalN造成的人胎肝细胞急性损伤具有保护作用。

Protective effect of granulocyte-colony stimulating factor on acute injury of human fetal hepatocytes

Objective To establish the model of acute injury of human fetal hepatocytes induced by D-galactosamine (D-GalN) and observe the protective effect of granulocyte-colony stimulating factor (G-CSF) on the injury of fetal hepatocytes. Methods The human fetal hepatocytes were hatched by gradient concentrations of D-GalN at different action time, and their activities were detected by MTT assay, in order to determine the optimal conditions for establishing the model of acute injury of human fetal hepatocytes. The human fetal hepatocytes were divided into 4 groups, which received different treatments: the first group (control group) was the blank control group; normal fetal hepatocytes of the second group (group G) were treated with G-CSF; the injury was induced by D-GalN in both the third group (group ND) and the fourth group (group GD), but the human fetal hepatocytes of group GD were treated with G-CSF, and those of group ND with the same amount of normal saline instead of G-CSF. The activities of the hepatocytes of each group were detected by both MTT assay and LDH release assay.Results More than 90% of human fetal hepatocytes could be killed by D-GalN at the concentration of 10 mg/ml for 12 hours. The medicine had enough time to play its protective effect. The activities of the hepatocytes were not significantly different between the control group and group G, while that of group GD was significantly higher than that of group ND (P<0.05).Conclusions The model of acute injury of human fetal hepatocytes can be established by D-GalN at the concentration of 10 mg/ml for 12 hours. G-CSF has a protective effect on the acute injury of human fetal hepatocytes induced by D-GalN.