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目的:探讨碱性成纤维细胞生长因子(bFGF)对间充质干细胞(MSCs)增殖的影响及其信号机制。方法:全骨髓贴壁法培养MSCs,流式细胞仪分析其表面MSCs标记(CD45、CD34、CD90、CD29)以及成骨、成脂肪等多向诱导分化潜能,鉴定其特征。MTT法分析不同浓度bFGF对MSCs增殖的影响,确立最佳浓度bFGF诱导MSCs增殖的时间特征,分别以磷脂酰肌醇3-激酶、促分裂素原活化蛋白激酶、磷脂酶C、蛋白激酶C、钙通道和钙泵选择性抑制剂等处理P3MSCs后,观察其对bFGF介导MSCs增殖的影响。结果:培养的MSCs表现CD90、CD29强阳性,具有成骨、成脂肪等多向分化能力;bFGF诱导MSCs增殖呈时间和剂量依赖特征,5.4×10-4mol/L bFGF作用48 h时MSCs增殖达峰值;磷脂酶C阻断剂U73122、钙通道阻断剂和选择性蛋白激酶C阻断剂等明显抑制了bFGF所介导的MSCs增殖效应;尤其是非选择性PKC抑制剂Straurosporine、钙泵选择性抑制剂thapsigargin以及蓝尼定受体抑制剂Ryandoine均抑制bFGF所诱导的MSCs增殖。结论:bFGF通过Ca2+/PKC/Erk1/2途径促进MSCs的增殖。
Objective: To investigate the effect of basic fibroblast growth factor (bFGF) on the proliferation of mesenchymal stem cells (MSCs) and its signal mechanism. Methods: MSCs were cultured with whole bone marrow adherent method. MSCs markers (CD45, CD34, CD90, CD29) on the surface were identified by flow cytometry. Differentiation and differentiation of MSCs were observed by flow cytometry. The effects of different concentrations of bFGF on the proliferation of MSCs were analyzed by MTT assay. The time characteristics of bFGF-induced MSCs proliferation were established. The effects of bFGF on the proliferation of MSCs were studied. After treatment of P3MSCs with calcium channel and selective inhibitors of calcium pump, the effects of bMSCs on the proliferation of MSCs were observed. Results: The cultured MSCs showed strong positive of CD90 and CD29, and had multipotential differentiation ability of osteoblast and adipocyte. The proliferation of MSCs induced by bFGF was time-and dose-dependent. MSCs proliferated up to 5.4 × 10-4 mol / L bFGF for 48 h Peak; Phospholipase C blockers U73122, calcium channel blockers and selective protein kinase C blockers significantly inhibited the bFGF-mediated proliferation of MSCs; especially non-selective PKC inhibitor Straurosporine, calcium pump selectivity Inhibitors of thapsigargin and Ryanidine receptor inhibitor Ryandoine inhibit bFGF-induced proliferation of MSCs. Conclusion: bFGF promotes the proliferation of MSCs via Ca2 + / PKC / Erk1 / 2 pathway.