基因修饰骨髓间充质干细胞脑源性神经营养因子的表达变化

摘要

目的：研究基因修饰的骨髓间充质干细胞( bone marrow mesenchymal stem cell,BMSC)分泌脑源性神经营养因子( brain-derived neurotrophic factor,BDNF)的表达变化。　　方法：实验分为空白对照组(未经转染BMSC细胞)、阴性对照组(采用不含BDNF基因的空载质粒转染的BMSC细胞)和实验组(采用含BDNF基因的质粒转染的BMSC细胞)。 Realtime PCR 测定基因修饰的 BMSC 细胞 BDNF mRNA表达,ELISA测定基因修饰的BMSC细胞BDNF的分泌表达。　　结果：实验组第3、4、5、6、7和8代 BMSC 细胞 BDNF mRNA表达高于空白对照组和阴性对照组,组间差异具有统计学意义( P3：F=491.788,P<0.05；P4：F=380.112,P<0.05；P5：F=1854.929,P<0.05；P6：F=224.540,P<0.05；P7：F=619.155,P<0.05；P8：F=10.092,P<0.05)。实验组BMSC细胞随着细胞传代,BDNF mRNA表达逐渐下降,不同代数间差异具有统计学意义(F=298.603,P<0.05)。实验组3、4、5、6、7和8代BMSC细胞BDNF的分泌表达高于空白对照组和阴性对照组,组间差异具有统计学意义(P3：F=520.609,P<0.05；P4：F=734.520,P<0.05；P5：F=152.847,P<0.05；P6：F=80.372,P<0.05；P7：F=96.083, P<0.05；P8：F=38.532,P<0.05)。实验组BMSC细胞随着细胞传代,BDNF的分泌表达逐渐下降,不同代数间差异具有统计学差异(F=230.084,P<0.05)。　　结论：通过基因工程可增强BMSC细胞表达BDNF。%AIM: To study the changes of brain - derived neurotrophic factor ( BDNF ) expression in gene modified bone marrow mesenchymal stem cells ( BMSC) . ●METHODS:BMSC were divided into blank control group ( without transfected BMSC ) , negative control group ( empty vector without BDNF gene transfected BMSC) and experimental group ( BDNF gene transfected BMSC) . The expression of BDNF mRNA in BMSC was measured by Realtime PCR, and the expression of BDNF in BMSC was measured by ELlSA. ●RESULTS:The BDNF mRNA expressions of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant (P3: F=491. 788, P<0. 05; P4: F=380. 112, P<0. 05;P5:F=1854. 929, P<0. 05; P6: F=224. 540, P<0. 05;P7:F=619. 155, P<0. 05; P8: F=10. 092, P<0. 05). As the BMSC cells in the experimental group passaging, the BDNF mRNA expressions in the experimental group decreased. The difference of BDNF mRNA expression among different passage cells was statistically significant (F=298. 603, P<0. 05). The BDNF secretion of 3, 4, 5, 6, 7 and 8-generation BMSC cells in the experimental group were higher than those in the blank control group and negative control group. The differences were statistically significant (P3: F=520. 609, P<0. 05; P4: F=734. 520, P<0. 05;P5:F=152. 847, P<0. 05;P6:F=80. 372, P<0. 05; P7:F=96. 083, P<0. 05;P8:F=38. 532, P<0. 05). As the BMSC cells in the experimental group passaging, the BDNF secretion decreased. The difference of BDNF secretion among different passage cells was statistically significant (F=230. 084, P<0. 05). ●CONCLUSION:Long-term expression of BDNF in BMSC can be enhanced by genetic engineering.