骨形态发生蛋白2在骨损伤不同时期的表达

目的:检测骨形态发生蛋白2(BMP2)在骨损伤不同时期的表达,了解其高峰表达时间。方法:实验组18只大鼠胫骨损伤造模,对照组6只大鼠假手术后处死,实验组平均分为3组,分别于术后第5天、第15天和第30天处死,提取大鼠骨组织总RNA,RT-PCR后扩增BMP2开放读框中编码成熟肽的基因片段,与pGEM-TEasy载体连接纯化后建立实时荧光定量PCR的标准曲线,检测BMP2的表达水平。结果:BMP2的表达在骨损伤后第5天开始升高,第15天达到高峰,第30天稍高于正常水平。结论:高峰表达时间的确定为进一步研究提供了理论上的依据。

Real Time PCR to Detect BMP2 Expression in Different Bone Deficiency Period

Objective : To study the peak-expression time of bone morphogenetic protein 2(BMP2) by detecting the expression of BMP2 in different period of bone deficiency. Methods: After tibia deficiency,the rats in control group were sacrificed immediately,and the 18 treated rats were divided into 3 groups,sacrificed on the 5th day,15th day and 30th day after operation. The total RNA from the bone of the rats was extracted. The encoding sequence gene of open reading frame was amplified from extracted total RNA by real time PCR(RT-PCR). The standard curve of RT-PCR was established with link to pGEM-T easy vector to detect the expressions of BMP2 in the experimental and control rats. Results: The expression of BMP2 started to raise on the 5th day,got to the peak level on the 15th day, and recovered to a slightly higher than normal level on the 30th day after bone deficiency. Conclusion: The peak expression time of BMP2 after bone deficiency provides a theoretical foundation for the further studies.