人参总皂苷增强成骨分化的间充质干细胞促造血作用研究

Total saponins of panax ginseng enhances the effect of osteoblast differentiation from mesenchymal stem cell on promoting hematopoiesis

目的观察人参总皂苷(TSPG)诱导间充质干细胞向成骨细胞分化的作用,探讨TSPG增强成骨分化的间充质干细胞促造血的可能机制。方法贴壁法培养人源骨髓间充质干细胞(h MSCs),成骨细胞分化和免疫表型进行鉴定。不同剂量的TSPG联合成骨细胞条件诱导液培养h MSCs,MTT法检测细胞增殖活性,对-硝基苯磷酸盐(p NPP)法检测培养上清碱性磷酸酶含量,茜素红染色法观察钙结节数量,Western blot法检测成骨细胞分化转录因子-核心结合蛋白因子2(RUNX2)蛋白表达水平,Elisa法检测培养上清造血相关因子含量,造血祖细胞集落生成实验检测培养上清造血支持能力。结果 MTT和p NPP结果均显示随着TSPG剂量增加,吸光值逐渐增加,呈剂量依赖性。钙结节数量和RUNX2蛋白表达水平明显高于对照组。造血相关生长因子和造血祖细胞集落数明显高于对照组。结论TSPG通过上调RUNX2蛋白表达诱导h MSCs向成骨细胞分化,同时增强成骨分化的MSCs支持造血。

Aim To observe mesenchymal stem cell differentiation into osteoblast induced by TSPG, and explore how TSPG enhances the promotion of hemato-poiesis of osteoblast differentiation from mesenchymal stem cell. Methods MSCs were cultured by TSPG combined with osteoinductive medium. The cellular vi-ability of proliferation was detected with MTT assay. The content of alkaline phosphatase in the cultural su-pernatant was tested with pNPP assay. The ability of MSCs to form calcium nodes was also observed after a-lizarin red stain. The protein expression of RUNX2 was analyzed with Western blot. The content of cytokines associated with hematopoiesis was tested with Elisa as-say. The ability of promoting hematopoiesis was detec-ted with hematopoietic colony forming assay. Results-nbsp;Both MTT and pNPP assay showed that optical den-sity （ OD） values were increased in response to TSPG treatment in a dose-dependent manner. The mineraliza-tion formation ability was enhanced with TSPG-treat-ment. Meanwhile, the expression of RUNX2 protein was up-regulated in TSPG-treated cell. Moreover, the content of cytokines associated with hematopoiesis and the number of hematopoietic progenitor colony were in-creased by TSPG-treatment compared with the control group. Conclusion TSPG could induce MSCs differ-entiation in to osteoblast and enhance the effect of oste-oblast differentiation from MSCs on promoting hemato-poiesis.