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目的探讨低频电磁场(LFEMF)对脑源性神经营养因子(BDNF)基因修饰的骨髓间充质干细胞(BMSCs)增殖的影响。方法通过同源重组的方法,用pAd-easy I腺病毒包装系统在体外构建含BDNF基因的重组腺病毒表达载体(rADBDNF);体外分离纯化SD大鼠来源的BMSCs,rADBDNF感染后,用LFEMF(50 Hz、5 mT)进行干预,连续干预3 d,并设对照组、BDNF基因修饰组、LFEMF组。在倒置相差显微镜下,对经台盼蓝染色的各组细胞进行计数;四甲基偶氮唑蓝(MTT)法对各组细胞的增殖效率进行检测;流式细胞仪分析细胞周期变化。结果 LFEMF作用于BDNF基因修饰后的BMSCs,细胞增殖速率与对照组、BDNF基因修饰组比较,差异有统计学意义(P<0.05),与LFEMF组相比,差异无统计学意义(P>0.05),G2/M期细胞数量明显增多。结论 LFEMF干预可促进BDNF基因修饰后BMSCs增殖。
Objective To investigate the effect of low frequency electromagnetic field (LFEMF) on the proliferation of bone marrow mesenchymal stem cells (BMSCs) modified by brain derived neurotrophic factor (BDNF) gene. Methods Recombinant adenovirus expression vector (rADBDNF) containing BDNF gene was constructed by homologous recombination in vitro using pAd-easy I adenovirus packaging system. BMSCs derived from SD rats were isolated and purified in vitro. After rADBDNF infection, 50 Hz, 5 mT) for 3 days. The control group, BDNF gene modified group and LFEMF group were given. Under inverted phase contrast microscope, the cells in each group stained with trypan blue were counted. The proliferation of each group was detected by MTT method. The cell cycle was analyzed by flow cytometry. Results LFEMF had a significant difference compared with control group and BDNF gene modified group (P <0.05), but there was no significant difference between LFEMF group and LFEMF group (P> 0.05) ), G2 / M phase cells increased significantly. Conclusion LFEMF intervention can promote the proliferation of BMSCs after BDNF gene modification.