成骨生长肽联合体外冲击波治疗对成骨细胞增殖功能的影响  

The effects of osteogenic growth peptide combined with extracorporeal shock waves on osteoblast proliferation

作　　者：钟俊[1] 刘世清[1] 魏金亮[1] 闫瑞承[1] 

机构地区：[1]武汉大学人民医院骨关节外科,武汉430061

出　　处：《中华物理医学与康复杂志》2012年第5期340-343,共4页Chinese Journal of Physical Medicine and Rehabilitation

基　　金：湖北省自然科学基金资助项目（2009CDB180）

摘　　要：目的探讨成骨生长肽（OGP）联合体外冲击波（ESW）干预对小鼠MC3T3-E1细胞增殖功能的影响。方法将传代培养的小鼠MC3T3-E1细胞分为对照组、实验组（给予OGP＋ESW干预）、ESW组（给予ESW干预）及OGP组（给予OGP干预）。于培养24h、48h及72h后采用MTT法进行细胞计数，于培养24h后采用倒置相差显微镜观察细胞增殖情况，并采用免疫组化法检测各组细胞蛋白激酶A（PKA）活性，于培养24h、48h后采用RT—PCR法检测PKAm-RNA表达。结果细胞计数及倒置相差显微镜观察结果均显示，实验组、ESW组及OGP组细胞增殖活性均显著强于对照组，并且实验组细胞增殖活性亦显著强于ESW组及OGP组；免疫组化检测结果显示，实验组、ESW组及OGP组PKA阳性染色明显强于对照组，并且实验组PKA阳性染色亦显著强于ESW组及OGP组；RT—PCR检测结果显示，实验组、ESW组及OGP组PKAm—RNA表达均明显强于对照组，并且实验组PKAm-RNA表达亦显著强于ESW组及OGP组。结论OGP与ESW联用具有协同作用，能进一步刺激MC3T3-E1细胞增殖与成熟。Objective To investigate the effects of osteogenic growth peptide （OGP） in combination with extracorporeal shock waves （ESWs） on osteoblast proliferation. Methods Passaged cells were divided into four groups for different treatments： a control group, an OGP ＋ ESW group, an ESW group, and an OGP group. After the respective treatments, the cells were cultured for 24 h, 48 h and 72 h and counted using methylthiazdy tetrazolium （MTT） and an inverted fluorescence microscope. Immunohistochemical examination was used for detecting protein kinase A （ PKA ） activity, and a reverse transcription-polymerase chair reaction （RT-PCR） was used for examining PKA mRNA expression at 24 and 48 hours. Results Cell counting revealed that cell proliferation in the OGP ＋ ESW, ESW and OGP groups was significantly promoted compared with the control group. Cell proliferation was greatest in the OGP ＋ ESW group. The immunohistochemical examination showed positive staining intensities in the OGP ＋ ESW, ESW and OGP groups significantly higher than in the control group. The positive staining intensity in the OGP ＋ ESW group was again the highest. PKA activity was also significantly higher in the OGP ＋ ESW, ESW and OGP groups than in the control group with the level in the OGP ＋ ESW group the highest. Conclusion OGP in combination with ESW has a synergistic effect in stimulating osteoblast proliferation and growth.

关 键 词：成骨生长肽 体外冲击波 蛋白激酶A MC3T3-E1细胞系 

分 类 号：R681[医药卫生—骨科学]