CAJ | 학술논문

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.
목적 탐토추화인자CCL2/수체CCR2(CCL2/CCR2)신호축삼여정기아이동동통적궤제.방법 선용200～ 300 g웅성SD대서,우상합좌측제일마아여절아간가력0.392N,근중이동상합마아.가력0(대조)、4h급1、3、5、7d,단백질인적법검측삼차신경절내추화인자CCL2급수체CCR2적단백표체(매개시간점3지대서);가력0(대조)、3、14d,면역조화법정위삼차신경절화삼차신경척속핵미측아핵(Vc핵)중CCL2양성세포(매개시간점3지대서).단백질인적법검측CCL2대체외배양적삼차신경원c-fos단백표체적영향(20지대서).분별사용CCL2중화항체、CCR2길항제후관찰대서적동물행위학변화(35지대서).결과 가력측삼차신경절내CCL2여CCR2단백표체수평균정현축점증가재명현하강적추세.CCL2표체수평재가력3、5、7d시(2.620±0.128、3.300±0.197、1.740±1.290)여대조(1.000±0.000)상비차이유통계학의의(P＜0.05),CCR2표체재가력3、5d시(1.636±0.061、1.766±0.126)여대조(1.000±0.000)차이유통계학의의(P＜0.05),이자봉치균출현재5d(분별증가위대조조적3.3화1.8배).CCL2주요표체우Vc핵천층성형효질세포,CCL2표체변화규률이급대서상해성안면정식행위규률여림상정기환자초시동통규률유사.외원성CCL2능명현격활삼차신경원,도치c-fos단백명현상조,CCR2길항제능유효억제대서상해성안면정식행위.결론 CCL2/CCR2신-호축적격활능조공정기아이동동통,재동통적발생、발전여유지중분연료중요적각색.
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 in TG neurons and astrocytes in Vc.Conclusions The CCL2/CCR2 axis was modulated by experimental tooth movement and involved in the development of tooth movement pain,and thus palyed an important role in orthodontic pain mechanism.