摘要
目的:探讨骨形态发生蛋白9(BMP9)与神经生长因子(NGF)联合处理对间充质干细胞(MSCs)骨向分化的影响。方法:以重组腺病毒法将BMP9导入C3H10T1/2细胞,构建MSCs骨向分化实验模型,设定分组为绿色荧光(GFP)对照组、NGF单独处理组、BMP9单独处理组以及BMP9+NGF联合处理组,分别在处理3 h、12 h、24 h、48 h、3 d、7 d后检测早期成骨分化标志物I型胶原、Runt相关转录因子2(RUNX2)和碱性磷酸酶(ALP)在细胞中的表达。检测方法采用ALP染色、实时定量RTPCR检测mRNA转录水平以及Western blot检测蛋白表达。结果:BMP9+NGF组较各单独处理组ALP活性显著增加,I型胶原、RUNX2的mRNA水平在BMP9+NGF组的表达明显高于其他各单独处理组,且各组在3 h即出现显著差异;RUNX2蛋白在各实验组均表达,BMP9+NGF组表达最高。结论:BMP9联合NGF在成骨诱导间充质干细胞分化早期,协同促进其骨向分化。
Objective： To investigate the effects of BMP9 combined with NGF on the osteogenic differentiation of mesenehymal stem cells（MSCs）. Methods： Recombinant BMP9 adenovirus was transfected into C3H10T1/2 cells. The cells were treated by CFP, NGF, BMP9 and BMP9 ＋ NGF respectively. The expression level of COL1, RUNX2 was detected by RT-PCR and Western blot, ALP activity was examined by ALP kit 3,12, 24,48 hours,3 and 7 days after treatment, respeetively. Results： The ALP activity of BMP9 ＋ NGF group was the highest among the 4 groups. The difference in the groups firstly appeared at 3 h after treatment. The highest expression level of RUNX2 and COL1 was detected in BMP9 ＋ NGF group. Conclusion ： NGF and BMP9 may synergisticly promote osteogenic dif- ferentiation at the early stage of osteogenic induction of C3H10T1/2 cells.
出处
《实用口腔医学杂志》
CAS
CSCD
北大核心
2017年第2期152-156,共5页
Journal of Practical Stomatology
基金
国家自然科学基金(编号:81371110)
