CAJ | 학술논문

目的通过建立选择性切断大鼠感觉/运动神经联合胫骨骨折的动物模型,研究感觉/运动神经损伤后对骨折愈合的影响,并检测骨折愈合过程中降钙素基因相关肽(calcitonin gene related peptide,CGRP)对骨保护素(osteoprotegerin,OPG)/破骨细胞分化因子(receptor activator nuclear factor kappa B ligand,RANKL)体系的影响,初步探讨周围神经调节骨折愈合的机制.方法取60只Wistar大鼠随机分为3组:前根(运动神经)切断+胫骨骨折组(anterior rhizotomy group,ART组)；后根(感觉神经)切断+胫骨骨折组(posterior rhizotomy group,PRT组)；单纯胫骨骨折组(sham operated group,SO组).分别于骨折术后7、14、21、28d处死大鼠,在骨折处上、下5mm部位取骨痂标本.免疫组织化学检测CGRP、OPG、RANKL的表达；采用软件Image-proplus 6.0进行图片分析；采用SPSS16.0进行统计学分析.结果 CGRP在SO组各时间点均呈强阳性表达,术后14 d和21 d,SO组、ART组和PRT组比较差异均有统计学意义(P＜0.05).OPG在SO组术后7d呈强阳性表达,以后逐渐下降但保持较高水平；术后14 d,PRT组和SO组、ART组比较差异有统计学意义(P＜0.05)；术后21 d,SO组和PRT组、ART组比较差异有统计学意义(P＜0.05).RANKL在SO组21 d为表达高峰,以后逐渐下降；术后14 d,PRT组和ART组、SO组比较差异有统计学意义(P＜0.05)；术后21 d,SO组和ART组、PRT组比较差异有统计学意义(P＜0.05).结论在骨折愈合过程中,感觉神经纤维对骨折愈合的影响较运动神经纤维显著.CGRP能够调节OPG/RANKL的表达量的比值,从而影响骨折的愈合过程.失神经支配(尤其是感觉神经)导致CGRP对OPG/RANKL表达的调节作用降低,这不利于骨折的愈合.完整的神经支配是正常骨折愈合的必要条件之一.
목적 통과건립선택성절단대서감각/운동신경연합경골골절적동물모형,연구감각/운동신경손상후대골절유합적영향,병검측골절유합과정중강개소기인상관태(calcitonin gene related peptide,CGRP)대골보호소(osteoprotegerin,OPG)/파골세포분화인자(receptor activator nuclear factor kappa B ligand,RANKL)체계적영향,초보탐토주위신경조절골절유합적궤제.방법 취60지Wistar대서수궤분위3조:전근(운동신경)절단+경골골절조(anterior rhizotomy group,ART조)；후근(감각신경)절단+경골골절조(posterior rhizotomy group,PRT조)；단순경골골절조(sham operated group,SO조).분별우골절술후7、14、21、28d처사대서,재골절처상、하5mm부위취골가표본.면역조직화학검측CGRP、OPG、RANKL적표체；채용연건Image-proplus 6.0진행도편분석；채용SPSS16.0진행통계학분석.결과 CGRP재SO조각시간점균정강양성표체,술후14 d화21 d,SO조、ART조화PRT조비교차이균유통계학의의(P＜0.05).OPG재SO조술후7d정강양성표체,이후축점하강단보지교고수평；술후14 d,PRT조화SO조、ART조비교차이유통계학의의(P＜0.05)；술후21 d,SO조화PRT조、ART조비교차이유통계학의의(P＜0.05).RANKL재SO조21 d위표체고봉,이후축점하강；술후14 d,PRT조화ART조、SO조비교차이유통계학의의(P＜0.05)；술후21 d,SO조화ART조、PRT조비교차이유통계학의의(P＜0.05).결론 재골절유합과정중,감각신경섬유대골절유합적영향교운동신경섬유현저.CGRP능구조절OPG/RANKL적표체량적비치,종이영향골절적유합과정.실신경지배(우기시감각신경)도치CGRP대OPG/RANKL표체적조절작용강저,저불리우골절적유합.완정적신경지배시정상골절유합적필요조건지일.
Objective To study the influence of sensory/motor nerve injury on fracture healing and the influence of calcitonin gene related peptide (CGRP) on osteoprotegerin (OPG)/receptor activator nuclear factor kappa B ligand (RANKL) system during the fracture healing process through the establishment of selective transection of sensory/motor nerve in rats with tibia fracture as an animal model and to elucidate the peripheral nerve injury regulation mechanism during fracture healing. Methods Sixty Wistar rats were divided into 3 groups randomly:motor nerve (ventral root) transection + tibia fracture group (anterior rhizotomy group,ART group) ; sensory nerve (dorsal root) transection + tibia fracture group (poeterior rhizotomy group,PRT group) ;simple tibia fracture group (sham operated group,SO group).The transverse tibia fracture was treated by intramedullary nailing.The rats were sacrificed at postoperative day 7,14,21 and 28 respectively to harvest the bony calluses 5 mm proximal and 5 mm distal to the fracture site.The expression of CGRP,OPG and RANKL was detected by immunohistochemistry and analyzed by Image-proplus 6.0 software.The comparison among the groups was analyzed using SPSS 16.0. Results Strong positive expression of CGRP was seen at each time point in SO group.At postoperative day 14 and 21,the differences of CGRP expression between SO group,ART group and PRT group were significant ( P ＜ 0.05).OPG expression was strongest at postoperative day 7 in SO group,while remaining high even with gradual decrease.The differences in OPG expression were significant ( P ＜ 0.05 ) at postoperative day 14 between PRT group and SO group and between PRT group and ART group,while that difference was seen between SO group and PRT group and between SO group and ART group at postoperative day 21.RANKL expression in the SO group peaked at postoperative day 21,and later gradually decreased.At postoperative day 14 the differences between PRT group and ART group and between PRT group and SO group were significant ( P ＜0.05),while at postoperative day 21 the differences between SO group and ART group and between SO group and PRT group were significant ( P ＜ 0.05). Conclusion During the fracture healing process,the influence of sensory nerve fibers was significantly higher than that of the motor nerve fibers.CGRP can regulate the OPG/RANKL expression ratio,thus affecting the healing process.Denervation (especially the sensory nerves) causes reduced CGRP regulation on OPG/RANKL expression,which is not favorable for fracture healing.Complete innervation is a necessary condition for the normal healing of fractures.