茵陈四苓颗粒对肝衰竭环境下骨髓间充质干细胞分化及IDO蛋白表达的影响

目的:研究肝衰竭体外模型环境下茵陈四苓颗粒对骨髓间充质干细胞的肝向分化及IDO蛋白表达的影响。方法:通过建立肝衰竭血清、中药干预血清、健康血清分别与骨髓间充质干细胞(BMSCs)共培养体系,检测BMSCs中CK18免疫荧光表达,Western-blot法检测甲胎蛋白(AFP)、肝组织白蛋白(ALB)、吲哚胺2,3—双加氧酶(IDO)蛋白表达。结果:1、CK18免疫荧光结果显示肝衰竭组及中药干预组染色均为弱阳性,但后者染色更强,而健康组无阳性染色;2、AFP、ALB在健康组、肝衰竭组、中药干预组均有表达,且各组间ALB比较均有差异(P0.05),三组AFP表达水平逐渐增高,各组间差异有统计学意义。IDO蛋白在健康组无表达,肝衰竭组表达低于中药干预组,且两组间差异具有统计学意义。结论:茵陈四苓颗粒能增强BMSCs在肝衰竭环境下的肝向分化趋势及免疫调节活性。

Effect of Yinchen Siling Granule on Differentiation and Expression of IDO Protein of Bone Marrow Mesenchymal Stem Cells in Hepatic Failure

ABSTRACT Objective: To study the effect of Yinchen Siling Granule on liver differentiation and IDO protein expression of bone marrow mesenchymal stem cells (BMSCs) in vitro model of hepatic failure. Methods: The co-culture system of hepatic failure serum, traditional Chinese medicine(TCM) serum and healthy serum with bone marrow mesenchymal stem cells (BMSCs) was established. The expression of CK18 in BMSCs was detected by immunofluorescence, and the expression of alpha-fetoprotein (AFP), hepatic albumin (ALB) and indoleamine 2-, 3-dioxygenase (IDO) protein were detected by Western-blot. Results: 1. The results of CK18 immunofluorescence showed that the staining of hepatic failure group and TCM group were weakly positive, and the latter staining was deeper. The healthy group had no positive staining; 2. The expressions of AFP, ALB were found in the healthy group, hepatic failure group and TCM group. There was significant difference in ALB expression among the three groups (P<0.05). The expression level of AFP in the three groups increased gradually, and there was significant difference among the three groups. There was no expression of IDO protein in the healthy group, but the expression in the hepatic failure group was lower than that in the TCM intervention group, and the difference between the two groups was statistically significant. Conclusion: Yinchen Siling Granule can enhance the liver differentiation trend and immunomodulatory activity of BMSCs in hepatic failure.