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目的 :观察糖尿病兔骨髓间充质干细胞(bone arrow-derived mesenchymal stem cells,BMSCs)以成骨分化为主的生物学特性。方法:通过静脉注射水合四氧嘧啶,建立糖尿病模型,以注射等量生理盐水作为对照组。在无菌条件下,局麻后抽取兔髂骨骨髓,获取BMSCs。采用全骨髓贴壁培养法对细胞进行纯化及培养,分别对2组细胞行MTT检测、实时定量PCR检测、碱性磷酸酶(ALP)活性检测,以及ERK、AKT信号通路表达检测。采用SPSS 13.0软件包对数据进行单因素方差分析(ANOVA)和SNK post hoc分析。结果:与正常组比较,实验组糖尿病兔BMSCs细胞增殖活性、成骨基因表达、ALP活性及成血管基因表达均显著下降(P<0.05)。结论:糖尿病兔BMSCs的增殖活性、成骨分化能力及成血管基因表达能力均受到损害。
Objective: To observe the biological characteristics of osteogenic differentiation of bone marrow-derived mesenchymal stem cells (BMSCs) in diabetic rats. Methods: Diabetic model was established by injecting hydrated alloxan intravenously, and the same amount of saline was injected as the control group. Under aseptic conditions, the rabbit iliac bone marrow was harvested to obtain BMSCs. The cells were purified and cultured by whole bone marrow adherent culture. MTT, real-time quantitative PCR, alkaline phosphatase (ALP) activity and ERK, AKT signaling pathway were detected in 2 groups of cells respectively. Data were subjected to one-way analysis of variance (ANOVA) and SNK post hoc analysis using SPSS 13.0 software package. Results: Compared with the normal group, the proliferative activity, osteogenic gene expression, ALP activity and angiogenic gene expression of BMSCs in experimental group were significantly decreased (P <0.05). Conclusion: The proliferative activity, osteogenic differentiation and angiogenesis of diabetic BMSCs are impaired.