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目的观察糖尿病大鼠模型骨髓间充质干细胞(BMSCs)体外增殖、抗凋亡和成骨分化能力。方法将40只6周龄雌性SD大鼠随机分为2组,每组20只,实验组腹腔注射链脲佐菌素,对照组注射等量的生理盐水。处理10周后,采用贴壁法获得2组大鼠的BMSCs。应用CCK-8检测第2代BMSCs增殖能力,0.3%过氧化氢和血清剥夺诱导的BMSCs凋亡;成骨诱导培养4周后,RT-PCR检测I型胶原蛋白(COL-I)、骨钙素(OCN)和核心结合蛋白因子-2(Runx-2)的mRNA表达水平;应用酶联免疫吸附法定量分析细胞碱性磷酸酶活性;茜素红染色及von Kossa染色分析矿化能力。结果链脲佐菌素成功诱导制备糖尿病大鼠模型。与对照组比较,实验组BMSCs增殖和抗凋亡能力降低,细胞成骨相关基因表达下降,细胞碱性磷酸酶活性显著降低,细胞成骨分化能力减弱(P<0.01)。结论糖尿病大鼠来源的BMSCs体外增殖、抗凋亡作用减弱,成骨分化能力显著下降。
Objective To observe the proliferation, anti-apoptotic and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in vitro in diabetic rats. Methods Forty six-week-old female Sprague-Dawley rats were randomly divided into two groups (20 rats in each group). The rats in the experimental group were injected intraperitoneally with streptozotocin and the control group with the same amount of saline. After 10 weeks of treatment, BMSCs of two groups of rats were obtained by adherence method. The proliferation of BMSCs was detected by CCK-8 and the apoptosis of BMSCs induced by 0.3% hydrogen peroxide and serum deprivation. After cultured for 4 weeks, the expression of COL-I, (OCN) and core-associated protein-2 (Runx-2). The activity of alkaline phosphatase was quantified by enzyme-linked immunosorbent assay. Alizarin red staining and von Kossa staining were used to analyze the mineralization ability. Results Streptozotocin successfully induced diabetic rat model. Compared with the control group, the proliferation and anti-apoptotic ability of BMSCs in the experimental group decreased, the expression of osteoblast-related genes decreased, the activity of alkaline phosphatase decreased significantly, and the ability of osteoblast differentiation decreased (P <0.01). Conclusion The proliferation and anti-apoptotic effects of BMSCs derived from diabetic rats decreased significantly, and the osteogenic differentiation ability decreased significantly.