CAJ | 학술논문

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( hMSCs),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hMSCs,MTT法检测细胞增殖活性,对-硝基苯磷酸盐( pNPP)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,West-ern 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pNPP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hMSCs向成骨细胞分化,同时增强成骨分化的MSCs支持造血。
목적：관찰인삼총조감( TSPG)유도간충질간세포향성골세포분화적작용,탐토TSPG증강성골분화적간충질간세포촉조혈적가능궤제。방법첩벽법배양인원골수간충질간세포( hMSCs),성골세포분화화면역표형진행감정。불동제량적TSPG연합성골세포조건유도액배양hMSCs,MTT법검측세포증식활성,대-초기분린산염( pNPP)법검측배양상청감성린산매함량,천소홍염색법관찰개결절수량,West-ern blot법검측성골세포분화전록인자-핵심결합단백인자2(RUNX2)단백표체수평,Elisa법검측배양상청조혈상관인자함량,조혈조세포집락생성실험검측배양상청조혈지지능력。결과 MTT화pNPP결과균현시수착TSPG제량증가,흡광치축점증가,정제량의뢰성。개결절수량화RUNX2단백표체수평명현고우대조조。조혈상관생장인자화조혈조세포집락수명현고우대조조。결론 TSPG 통과상조RUNX2단백표체유도hMSCs향성골세포분화,동시증강성골분화적MSCs지지조혈。
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 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.