柔肝化纤颗粒促进BMSC向肝脏归巢及其机制研究

Mechanism of Rougan Huaxian Granules Promoting BMSC Homing to Liver

目的研究柔肝化纤颗粒促进骨髓来源间充质干细胞(BMSC)向肝脏归巢的作用及其机制。方法动物实验部分,大鼠分为肝纤维化模型加BMSC移植组,及肝纤维化模型加BMSC移植联合柔肝化纤干预组,BMSC细胞实验分为对照组,柔肝化纤含药血清(低、中、高浓度)干预组。分别采用qPCR,WesternBlot法检测SDF-1、CD62P、CD62E、CD44、CXCR4、VCAM-1和CD49D mRNA及蛋白表达情况,冷冻切片检测携带绿色荧光蛋白的BMSC向肝组织归巢情况。结果与单独BMSC移植组相比,柔肝化纤颗粒联合干细胞干预组大鼠肝脏干细胞粘附相关基因和干细胞归巢相关基因mRNA及蛋白表达量显著上调(P<0.05),与单独BMSC移植组相比,柔肝化纤颗粒联合干预组大鼠肝脏内携带绿色荧光蛋白的BMSC数量明显增加。细胞实验部分,与对照组相比,添加高浓度柔肝化纤含药血清能够显著上调干细胞归巢相关基因mRNA及蛋白表达量(P<0.05)。结论柔肝化纤颗粒能够促进BMSC向纤维化肝组织归巢,并能够提高肝脏募集干细胞聚集水平,其作用机制可能与SDF-1/CXCR4轴有关。

Objective To study the effect and mechanism of Rougan Huaxian Granuleson homing of bone marrowderived mesenchymal stem cells(BMSC)to the liver.Methods In the animal experiment,the rats were divided into liver fibrosis model plus BMSC transplantation group,and liver fibrosis model plus BMSC transplantation combined with soft liver chemical fiber intervention group.The BMSC cell experiment was divided into control group,and Rougan Huaxian Granuleson containing serum(low,medium and high concentration)intervention group.MRNA and protein expressions of sdf-1,CD62 P,CD62 E,CD44,CXCR4,vcam-1 and CD49 D were detected by qPCR and WesternBlot respectively,and the homing of BMSC with green fluorescent protein to liver tissue was detected by frozen section.Results Compared with the single BMSC transplantation group,liver fiber particles combined stem cells in the intervention group in the rat liver stem cell adhesion related gene and stem cell homing related genes mRNA and protein expression quantity rise significantly(P<0.05),compared with the single BMSC transplantation group,liver fiber particles combined intervention group in the liver of rats with green fluorescent protein significantly increased the number of BMSC.In the cell experiment,compared with the control group,the mRNA and protein expression levels of homing-related genes of stem cells were significantly up-regulated by the addition of high-concentration serum containing livers(P<0.05).Conclusion The Rougan Huaxian Granuleson can promote the homing of BMSC to fibrosis liver tissue and improve the aggregation level of stem cells in liver recruitment.The mechanism may be related to sdf-1/CXCR4 axis.