一贯煎促进骨髓间充质干细胞逆转肝纤维化的实验研究

The promotion of Yiguanjian on the recovery of liver fibrosis through mesenchymal stem cells: an experimental study

目的该研究观察骨髓间充质干细胞对肝纤维化的逆转作用,了解一贯煎对该过程的作用,阐释一贯煎逆转肝纤维化的机制。方法 60只雄性Sprague-Dawlay大鼠随机分为5组,即正常对照组、模型组、阳性对照组、骨髓间充质干细胞组、一贯煎加骨髓间充质干细胞组,每组12只。除正常对照组外,均建立肝纤维化恢复模型,分离、培养和标记骨髓间充质干细胞,在CCl4停止注射后3天,骨髓间充质干细胞组每只动物经尾静脉注射5×106mmol/L浓度1 mol骨髓间充质干细胞;一贯煎加骨髓间充质干细胞组每只大鼠尾静脉注射等量的骨髓间充质干细胞外,给予一贯煎浓缩液2 ml/200 g灌胃,每天1次。阳性对照组给予秋水仙碱灌胃,0.25 mg/kg,每天1次。正常对照组和模型组按相同方法灌服等量生理盐水。阳性对照组、正常对照组和模型组经尾静脉注射等量生理盐水。4周后,检测各组大鼠血清谷丙转氨酶、谷草转氨酶等肝功能指标;通过HE染色评价肝脏炎症活动度(G)和纤维化程度(S);荧光显微镜下示踪肝组织中骨髓间充质干细胞。结果 (1)骨髓间充质干细胞组和一贯煎加骨髓间充质干细胞组较模型组和阳性组在改善肝功能、减轻炎症、逆转肝纤维化的作用较好,P<0.05。一贯煎加骨髓间充质干细胞组较骨髓间充质干细胞组在改善肝功能、逆转纤维化的作用更明显,P<0.05。(2)骨髓间充质干细胞组和一贯煎加骨髓间充质干细胞组骨髓间充质干细胞在肝脏分布面积较模型组增大,P<0.05;一贯煎加骨髓间充质干细胞组较其他组差异显著,P<0.05。结论骨髓间充质干细胞具有逆转肝纤维化的作用,一贯煎对该过程具有促进作用;一贯煎能够促进骨髓间充质干细胞募集到肝脏,从而发挥逆转肝纤维化的作用。

Objective To investigate the effect of bone marrow mesenchymal stem cells on liver fi-brosis reversal, understand the role of Yiguanjian on this process, and interpret the mechanism of Yiguan-jian on the recovery of liver fibrosis. Methods we established liver fibrosis rat model. 60 male SD rats were divided into 5 groups：Normal group, model group, western medicine group, mesenchymal stem cells group and Yiguanjian combined with bone marrow mesenchymal stem cells group, There were 12 rats in each group. Except the normal group, hepatic fibrosis model was established. Isolation, culture and la-beled bone marrow mesenchymal stem cells. On the third day after CCl4 injection, each rat in mesenchy-mal stem cells group was given intravenous injection of 5 × 10^6 mmol/L 1 mol bone marrow mesenchymal stem cells. Except equal amounts of bone marrow mesenchymal stem cells, each rat in Yiguanjian com-bined with bone marrow mesenchymal stem cells group was given Yiguanjian（2 ml/200 g d）by intragastric administration. Western medicine group was given colchicine （0. 25 mg/kg d） by intragastric administra-tion. The normal group and model group were fed with same volume of saline daily. Western medicine group, normal group and model group was given intravenous injection of same volume of saline daily. After 4 weeks,the serum ALT and AST were tested, the HE staining were used to evaluate the degree of liver in-flammation and fibrosis （ G） （ S） , Bone marrow mesenchymal stem cells in liver tissue were observed under fluorescence microscop. Results Compared with the model group and western medicine group,mesenchymal stem cells group and Yiguanjian combined with bone marrow mesenchymal stem cells group had better effect in the improvement of liver function, reduce inflammation, reversing hepatic fibrosis, P 〈 0. 05. Compared with the mesenchymal stem cells group , Yiguanjian combined with bone marrow mesenchymal stem cells group had better effect on improving the liver function, reversing hepatic fibrosis, P 〈 0. 05. Compared with the model group, the area of bone marrow mesenchymal stem cells in liver was greater in mesenchymal stem cells group and Yiguanjian combined with bone marrow mesenchymal stem cells group, P 〈 0. 05. Compared with the other groups, the area of bone marrow mesenchymal stem cells in liver was greater in Yiguanjian combined with bone marrow mesenchymal stem cells group, P〈 0. 05. Conclusion Bone marrow mesen-chymal stem cells can reverse liver fibrosis , Yiguanjian can promote this process, Yiguanjian can promote bone marrow mesenchymal stem cell recruitment to the liver, thus play an effect of recovery of liver fibrosis.