摘要
目的探讨骨形态发生蛋白-7(BMP-7)联合胰岛素样生长因子-1(IGF-1)对体外培养兔退变髓核细胞(NPCs)生物学活性的影响。方法通过建立兔腰椎间盘退变模型及体外培养兔退变NPCs,将实验兔随机分为4组,即对照组、BMP-7干预组、IGF-1干预组、BMP-7和IGF-1联合干预组。BMP-7和IGF-1干预3d、5 d、7 d后,通过MTT法检测各组细胞活力,用流式细胞术检测各组细胞凋亡情况,RT-PCR检测NPCs中Ⅱ型胶原和蛋白聚糖(PG)的mRNA水平,ELISA检测各组细胞液中Ⅱ型胶原蛋白含量,二甲基亚甲基蓝比色法检测各组PG的含量。结果 BMP-7或IGF-1单独或联合给药都能显著提高NPCs活力、抑制细胞凋亡,并提高Ⅱ型胶原和PG mRNA及蛋白含量,且联合给药促进程度大于单独给药,差异有统计学意义(P<0.05)。结论 BMP-7联合IGF-1可提高体外培养兔退变NPCs活力及功能性蛋白的表达。
Objective To explore the effect of bone morphogenetic protein( BMP- 7) and insulin- like growth factors- 1( IGF- 1) on the bioactivityl of the rabbit degenerative nucleus pulposus cells( NPCs). Method Nucleus pulposus cells were isolated from rabbit lumbar intervertebral disc degeneration model. The cells was divided into five groups: control group,BMP- 7 intervention group,IGF- 1 intervention group and BMP- 7 plus IGF- 1 intervention group. After intervented by BMP- 7 and IGF- 1 NPCs for 3 d,5 d and 7 d,the viability was measured by MTT method. Flow cytometry was used to detect the cell apoptosis. The level of collagen Ⅱand aggrecan( PG) mRNA was assayed by RT- PCR. The level of collagen Ⅱ protein was assayed by ELISA method. The level of PG protein was measured by 1,9- dimethyl methylene blue technology. Results BMP- 7 and IGF- 1 could not only increase the viability of NPCs cell and inhibit the apoptosis,but also increase the expression of BMP- 7 and IGF- 1mRNA and protein. Moreover,the effect of joint administration was better than that of administrated alone,the difference was significant( P < 0. 05). Conclusion BMP- 7 combined with IGF- 1 could increase the viability of NPCs and promote the expression of functional protein in NPCs.
出处
《宁夏医学杂志》
CAS
2016年第5期448-451,共4页
Ningxia Medical Journal
基金
宁夏自然科学基金资助项目(NZ13183)
