人基质细胞衍生因子1对骨髓间充质干细胞增殖的影响.doc

人基质细胞衍生因子 1对骨髓间充质干细胞增殖的影响  姓 名： 2014年6月25日 人基质细胞衍生因子1对骨髓间充质干细胞增殖的影响 ：孔霞，唐俊明，郭凌郧，杨建业，陈龙，黄永章，王家宁 目的：探索人基质细胞衍生因子1(SDF 1)对间充质干细胞(MSC) 增殖的影响.方法：采用经典的全骨髓贴壁法培养MSC，通过成骨、成脂肪等多 向诱导分化以及流式细胞仪分析其表面标记(CD133, CD34, CD90, CD105) 等鉴定MSC特征；以P3 MSC为实验材料，采用H3 TdR参入法分析SDF 1 对 MSC 增殖的影响.以 50 nmol/L 渥曼青霉素，10 mmol/L LY294002, 50 mmol/L PD98059, 0.1g/LAMD3100 等分别处理 P3 MSC,观察 SDF 1 影响 MSC ±汽殖 的信号机制.结果：培养的MSC呈现出CXCR4, CD90和CD105强阳性，具有 成骨、成脂肪等多向分化能力；含有100 mg/L SDF 1的20 mL/L促进了 MSC 增殖，而含有100 mg/L SDF 1的150 mL/L FBS抑制了 MSC增殖.SDF 1抑 制MSC增殖的效应与其使MSC停滞在G1/S期有关.促分裂原活化蛋白激酶 P38MAPK阻断剂SB203580和CXCR4阻断剂AMD3100取消了 SDF 1抑制 MSC增殖的效应，而细胞外信号调节激酶阻断剂PD98059加强了 SDF 1抑制 MSC增殖的效应.结论：SDF 1/CXCR4介导的抑制MSC增殖的效应与促分裂 原活化蛋白激酶和细胞外信号调节激酶等信号分子有关. 关键词 人基质细胞源衍生因子1;间充质干细胞;增殖;促分裂原活化蛋 白激酶 Abstract AIM: To explore the effect of stromal cell derived factor 1 alpha (SDF-la) on mesenchymal stem cell (MSC) proliferation. METHODS: MSC culture was performed with the classical whole bone marrow adhering method; MSC was identified through multi differentiation and surface marker assay (CXCR4, CD 133, CD34, CD90，CD105); After MSCs were treated with 100 mg/L SDF la, 50 nmol/L wortmannin, 10 mmol/L LY294002, 50 mmol/L PD98059 and 0.1 g/L AMD3100 inhibitors, the proliferation rates were measured by [3H] thymidine incorporation assay. RESULTS: Cultured MSCs had the ability of multi differentiation and highly expressed CXCR4, CD 105 and CD90. 20 mL/L FBS including 100 mg/L SDF la promoted MSC proliferation，however exposure of MSCs to 150 mL/L FBS including 100 mg/L SDF la could induce marked inhibition of MSC proliferation through the arrest in the Gl/S checkpoint. And its inhibitory effect on MSC proliferation was partly reversed by p38 mitogen activated protein kinase (MAPK) inhibitor SB203580 and CXCR4 inhibitor AMD3100. However its inhibition effect on MSC proliferation was obviously enhanced by extracellular signal regulated kinase (ERK1/2) inhibitor PD98059.CONCLUSION: The effect of SDF la on MSC proliferation is related to ER