不同时机转染基因对下颌骨牵引区细胞周期蛋白表达的影响研究  

Effect of gene transfecting at different time on expression of cyclins in mandibular distraction area

作　　者：尹康 胡纯兵[2] 周滨[2] 吴国平[2] 赵利平[3] 

机构地区：[1]西南医科大学附属医院外科,四川泸州646000 [2]济宁医学院附属医院烧伤整形科,山东济宁272029 [3]南京医科大学附属友谊整形外科医院,南京210029

出　　处：《重庆医学》2017年第20期2737-2740,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(30600653);四川省卫生厅科研课题(12226)

摘　　要：目的通过观察不同时机转染基因对下颌骨牵引区细胞周期蛋白表达的影响,探讨基因治疗促进牵引成骨的分子机制及最佳时机。方法 48只新西兰大白兔建立双侧下颌骨牵引成骨模型后均分为A、B、C、D组。A、B、C组分别于术后即刻、术后第4天、术后第14天在牵引区转染重组质粒pIRES-hBMP2-hVEGF165,并局部给予电穿孔刺激。4组动物均在术后第4天,以1mm/d的速度持续牵引10d后进入固定期。于固定期第7、14、28、56天各组分别处死动物3只,通过免疫组织化学染色检查牵引区新生骨组织cyclin A、D1、E的表达。结果在固定期第7、14天时,A、B、C组牵引间隙cyclin A、D1、E的表达较强,在第7天时表达最强烈,并且A、B、C组均较D组表达强;第28、56天时各组表达均较弱。在第7天时,B组表达较A、C、D组强(P<0.05),A、C组间表达差异无统计学意义(P>0.05),但均较D组强(P<0.05);在第14天时,B、C组表达较A、D组强(P<0.05),B、C组间及A、D组间表达差异无统计学意义(P>0.05)。结论 cyclin A、D1、E的高表达能促进牵引区细胞分裂、增殖和分化,进而加快牵引区新骨形成,牵引期是基因治疗干预下颌骨牵引成骨的最佳时机。Objective To investigate the optimal time and molecular mechanism of gene therapy for promoting distraction os- teogenesis by observing the effect of gene transfecting at different time on expression of cyclins in mandibular distraction area. Methods Forty eight New-Zealand rabbits were employed. After accomplishing bilateral mandibular distraction osteogenesis mod- el, the rabbits were randomly divided into the group A,B, C and D. The group A, B and C were transfected by recombinant plasmids pIRES-hBMP2-hVEGF165 2 μg（0.1 μg/μL）at the distraction area instantly after operation, on postoperative 4,14 d and given local electroporation stimulation. The four groups entered the consolidation stage after 10 d continuous traction at a rate of 1 mm/d on postoperative 4 d. Three rabbits in each group were respectively sacrificed on 7,14,28,56 d of the consolidation stage. The expres- sion of cyclin A,Dl,and E of fresh bone tissue in distraction area were detected by immunohistochemical staining. Results Cyclin A, D1, and E were strongly expressed in the traction gap on 7,14 d of consolidation stage found, which was strongest on 7 d, moreo- ver the group A,B and C were stronger than the group D. The expression in each group was weakened on 28,56 d. The expression on 7 d in the group B was stronger than that in the4 group A,C and D（P〈0. 05）,the expression had no statistical difference be- tween the group A and C（P^0.05） ,but all were stronger than the group D（P〈0.05） the expression in the group B and C on 14 d was stronger than that in the group A and D,but the expression had no statistical difference between the group B and C and be- tween the group A and D（P〈0.05）. Conclusion The high expression of cyclin A,D1 and E can promote the cellular division,pro- liferation and differentiation in distraction area, thus accelerates the new bone formation in the distraction area, prompting that the distraction period is the best time for distraction osteogenesis under gene therapeutic intervention.

关 键 词：转染时机 电穿孔 基因疗法 牵引成骨 细胞周期蛋白 

分 类 号：R782[医药卫生—口腔医学]