人肝源性干细胞腹腔移植抗刀豆蛋白A诱导小鼠急性肝损伤的机制研究

The mechanisms of intraperitoneal transplantation of human liver-derived stem cells against concanavalin A-induced acute liver injury in mice

目的探讨人肝源性干细胞抗刀豆蛋白A(ConA)诱导的小鼠急性肝损伤的作用机制。方法将72只雄性C57BL/6小鼠分成2组,ConA模型组和干细胞干预组,干预组腹腔注射人肝源性干细胞,模型组注射等量的磷酸盐缓冲液,干预12h后尾静脉注射ConA,注射ConA后分5个不同时间点(0h、3h、6h、12h、24h)分别处死小鼠,检测血清ALT、AST、TNF-α和IL-10的水平,检测肝组织TNF-α和IL-10的mRNA表达水平,观察肝组织病理改变及细胞凋亡程度。结果干预组小鼠(12h、24h)较模型组相比血清ALT、AST水平显著下降(P<0.05),肝组织坏死程度明显减轻;干预组小鼠(3h、6h、12h)血清TNF-α水平较模型组明显下降(P<0.05),而血清IL-10(6h)水平较模型组明显升高(P<0.05);肝组织TNF-α和IL-10的mRNA水平变化与血清基本一致;TUNEL染色显示干预组小鼠(12h、24h)肝组织凋亡程度较模型组明显减轻。结论人肝源性干细胞抑制促炎因子释放、促进抑炎因子表达及抗凋亡作用可能是其保护ConA诱导小鼠急性肝损伤的重要机制。

Objective To investigate the mechanisms of human liver-derived stem cells against concanavalin A（ConA）-induced acute liver injury in mice.Methods A totle of 72 male C57 BL/6 mice were randomly divided into 2 groups,the ConA model group and the stem cell intervention group.The intervention group was injected intraperitoneally with human hepatic stem cells and the model group was injected with the same amount of phosphate buffer.After12 hof intervention,ConA was injected into the tail vein and mice were sacrificed at 5 different time points（0 h,3 h,6 h,12 h,and 24 h）.The levels of serum ALT,AST,TNF-αand IL-10 were detected,and the expressions of mRNA of TNF-αand IL-10 in liver tissues were measured.The pathological changes and apoptosis of liver tissues were observed.Results Compared with the ConA model group,the levels of serum ALT and AST in the intervention group（12 h,24 h）were significantly decreased（P〈0.05）,and the degree of hepatic tissue necrosis was significantly reduced.The levels of serum TNF-αin the intervention group（3 h,6 h,12 h）were significantly lower than those in the model group（P〈0.05）,while the levels of serum IL-10（6 h）were significantly higher than those in the model group（P〈0.05）.The levels of mRNA of TNF-αand IL-10 in liver tissue were basically consistent with those in serum.TUNEL staining showed that the degree of liver cell apoptosis in the intervention group mice（12 h,24 h）was significantly reduced compared with the model group.Conclusion Human liver-derived stem cells protects against ConA-induced liver injury,which may be mediated by inhibiting the release of pro-inflammatory factors,promoting the expression of anti-inflammatory cytokines and anti-apoptosis.