趋化因子CCL2/受体CCR2信号轴激活调控正畸牙移动疼痛的实验研究  

CCL2/CCR2 signaling activation contributes to tooth movement pain

作　　者：杨秩[1,2,3] 罗薇 傅润卿[1,2,3] 谭宇[1,2,3] 袁玲君[1,2,3] 房兵[1,2,3] 

机构地区：[1]上海交通大学医学院附属第九人民医院口腔医学院口腔颅颌面科,200011 [2]上海市口腔医学研究所,200011 [3]上海市口腔医学重点实验室,200011 [4]上海市口腔病防治院正畸科

出　　处：《中华口腔医学杂志》2014年第8期500-505,共6页Chinese Journal of Stomatology

基　　金：国家自然科学基金（81000451）;上海市自然基金（14ZR1424100）;上海市卫生局青年科研项目（2010Y142）;上海高校选拔培养优秀青年教师科研专项基金（2010）

摘　　要：目的 探讨趋化因子CCL2/受体CCR2（CCL2/CCR2）信号轴参与正畸牙移动疼痛的机制.方法 选用200～ 300 g雄性SD大鼠,于上颌左侧第一磨牙与切牙间加力0.392N,近中移动上颌磨牙.加力0（对照）、4h及1、3、5、7d,蛋白质印迹法检测三叉神经节内趋化因子CCL2及受体CCR2的蛋白表达（每个时间点3只大鼠）;加力0（对照）、3、14d,免疫组化法定位三叉神经节和三叉神经脊束核尾侧亚核（Vc核）中CCL2阳性细胞（每个时间点3只大鼠）.蛋白质印迹法检测CCL2对体外培养的三叉神经元c-fos蛋白表达的影响（20只大鼠）.分别使用CCL2中和抗体、CCR2拮抗剂后观察大鼠的动物行为学变化（35只大鼠）.结果 加力侧三叉神经节内CCL2与CCR2蛋白表达水平均呈现逐渐增加再明显下降的趋势.CCL2表达水平在加力3、5、7d时（2.620±0.128、3.300±0.197、1.740±1.290）与对照（1.000±0.000）相比差异有统计学意义（P＜0.05）,CCR2表达在加力3、5d时（1.636±0.061、1.766±0.126）与对照（1.000±0.000）差异有统计学意义（P＜0.05）,二者峰值均出现在5d（分别增加为对照组的3.3和1.8倍）.CCL2主要表达于Vc核浅层星形胶质细胞,CCL2表达变化规律以及大鼠伤害性颜面整饰行为规律与临床正畸患者初始疼痛规律类似.外源性CCL2能明显激活三叉神经元,导致c-fos蛋白明显上调,CCR2拮抗剂能有效抑制大鼠伤害性颜面整饰行为.结论 CCL2/CCR2信-号轴的激活能调控正畸牙移动疼痛,在疼痛的发生、发展与维持中扮演了重要的角色.Objective To test the hypothesis that the CCL2/CCR2 signaling pathway plays an important role in pain induced by experimental tooth movement.Methods Male Sprague-Dawley rats weighing between 200 and 300 g were used in this study.Expression of CCL2/CCR2 in the trigeminal ganglion（TG） was determined by Western blotting 0 h,4 h,1 d,3 d,5 d,7 d after tooth movement.Localization of the CCL2 was revealed by immunohistochemistry.Changes in body weight,nocifensive behaviors,and the effects of CCL2/CCR2 antagonists on these changes in pain behaviors were evaluated.Exogenous CCL2 was injected into periodontal tissues and added to TG neurons in culture and the resulting c-fos expression and pain responses were detected.In addition,the expression and cellular localization of CCL2 in the medullary dorsal horn （MDH） was determined by immunohistochemistry 3 d and 14 d after tooth movement.Results Experimental tooth movement led to a statistically significant increase in CCL2/CCR2 expression at the protein level from day 3 to 7 after application of force initiating tooth movement.When compared with control group（1.000± 0.000）,CCL2 increased to （2.620 ± 0.128）,（3.300±0.197） and （1.740±1.290） at day 3,5 and 7 respectively,which were statistically significant （P〈0.05）.CCR2 expression levels were （1.636±0.061） and （1.766±0.126） compared with that in control group （1.000±0.000） at day 3 and 5 respectively with statistical significance （P〈0.05）.Both of them peaked on day 5 （3.3 and 1.8 time compared to control group）.Application of recombinant CCL2 led to the up-regulation of c-fos expression in vivo and in vitro,and triggered a corresponding nocifensive behavior in rats.The magnitude of the nocifensive behavior could be reduced by a CCR2 antagonist,and by CCL2 neutralizing antibody.Furthermore,we found a significant increase in the expression of CCL2,corresponding well to the upregulation of the time spent on nocifensive behaviors after ETM.In addition,CCL2 was up-regulated i

关 键 词：趋化因子CCL2 受体 CCR2 牙移动 信号传导 

分 类 号：R783.5[医药卫生—口腔医学]