摘要目的 观察在激素诱导下,人骨髓间充质干细胞(hBMSCs)内降钙素基因相关肽受体(CGRPR)、P物质受体(SPR)、过氧化物酶体增殖子活化受体γ(PPARγ)、成骨转录因子Runx2以及B淋巴细胞/白血病-2(bcl-2)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)等基因表达的变化.方法 密度梯度离心和贴壁培养获得hBMSCs,流式细胞仪鉴定后,将传代培养的第3代细胞随机分为两组,实验组加入浓度为l×10-7 mmol/L的地塞米松,对照组正常培养.于诱导后的第7天,实时定量-聚合酶链反应(RT-qPCR)分别测定bcl-2 mRNA、Caspase-3 mRNA、CGRPR mRNA、SPR mRNA、Runx2 mRNA以及PPARγmRNA的表达.结果 分离培养的hBMSCs呈均匀一致的长梭形,排列成旋涡状或放射状.实验组Caspase-3 mRNA、PPARγ mRNA较对照组明显升高,2-△△Ct值分别为5.99±2.00、6.93±0.70;而bcl-2 mRNA、CGRPR mRNA、SPR mRNA、Runx2 mRNA较对照组明显降低,2-△△Ct值分别为0.20 ±O.51、0.11±0.02、0.28±0.08、0.13±0.21,差异均有统计学意义(P＜0.01).结论 激素能够上调hBMSCs中凋亡基因、成脂基因表达,下调抗凋亡基因、神经肽因子、成骨基因表达,促进细胞凋亡、成脂分化,抑制其成骨分化.

abstractsObjective To observe the effect of steroids on the expression of calcitonin gene-related peptide receptor (CGRPR),substance P receptor (SPR),perxisome proliferators activated receptor-γ (PPARγ),Runx2,B lymphocytes/leukemia-2 (bcl-2) and Caspase-3 in human bone marrow mesenchymal stem cells (hBMSCs).Methods hBMSCs were isolated and purified by density gradient centrifugation combined with plastic-adhering culture,and identified by flow cytometry.The hBMSCs of the 3rd generation were randomly divided into two groups.The control group received no treatment,while cells in the experimental group were treated with 1 × 10-7 mol/L dexamethasone.The mRNA was isolated and the expression level of bcl-2,Caspase-3,CGRPR,SPR,Runx2 and PPARγ was detected by using RT-qPCR on the 7th day after treatment.Results After isolation and culture,the hBMSCs displayed a distinct phenotype-the spindle-shaped,and uniform cells grew into clusters with swirl or radiation pattern.The expression levels of Caspase-3 and PPARγ in the experimental group were significantly higher than those in the control group,with the 2-△△Ct being 5.99 ±2.00 and 6.93 ±0.70 respectively (P ＜0.01).The expression levels of bcl-2,CGRPR,SPR and Runx2 were lower in the experimental group than those in the control group,with the 2-△△Ct being 0.20 ±0.51,0.11 ±0.02,0.28 ±0.08 and 0.13 ±0.21 respectively (P ＜0.01).Conclusion The steroid can up-regulated the expression of apoptosis gene and adipogenic gene in hBMSCs,and down-regulated the expression of anti-apoptosis gene and neuropeptide factors,which can promote the apoptosis and adipogeneic differetiantion of the cells and inhibit their osteogenic differentiantion.