CAJ | 학술논문

目的评价脊髓趋化因子CXC配体13(CXCL13)在大鼠神经病理性痛中的作用.方法雄性成年SD大鼠108只,体重150～200g,采用随机数字表法,将其分为4组(n=27):假手术组(S组)、神经病理性痛组(NP组)、小干扰RNA(siRNA)阴性对照(NC-siRNA)组(NS组)和CXCL13-siRNA组(CS组).NP组、NS组和CS组采用结扎L5脊神经的方法制备大鼠神经病理性痛模型,S组仅暴露L5脊神经,但不结扎.NS组和CS组分别鞘内注射NC-siRNA慢病毒和CXCL13-siRNA慢病毒10μl.分别于术后3、7和14 d时测定机械痛阈,然后处死大鼠,取脊髓组织,采用免疫荧光法检测CXCL13和Neun的共表达情况以及星形胶质细胞活化情况.采用Western blot法测定CXCL13和GFAP的蛋白表达,采用RT-PCR法测定CXCL13和GFAP的mRNA表达.结果与S组比较,NP、NS组和CS组术后各时点机械痛阈下降,脊髓CXCL13和GFAP的蛋白及其mRNA表达上调(P＜0.05)；与NP组比较,CS组术后各时点机械痛阈升高,脊髓CXCL13和GFAP的蛋白及其mRNA表达下调(P＜0.05),NS组各时点机械痛阈、脊髓CXCL13和GFAP的蛋白及其mRNA表达差异无统计学意义(P＞0.05).结论脊髓CXCL13通过激活星形胶质细胞参与大鼠神经病理性痛的形成和维持.
목적 평개척수추화인자CXC배체13(CXCL13)재대서신경병이성통중적작용.방법 웅성성년SD대서108지,체중150～200g,채용수궤수자표법,장기분위4조(n=27):가수술조(S조)、신경병이성통조(NP조)、소간우RNA(siRNA)음성대조(NC-siRNA)조(NS조)화CXCL13-siRNA조(CS조).NP조、NS조화CS조채용결찰L5척신경적방법제비대서신경병이성통모형,S조부폭로L5척신경,단불결찰.NS조화CS조분별초내주사NC-siRNA만병독화CXCL13-siRNA만병독10μl.분별우술후3、7화14 d시측정궤계통역,연후처사대서,취척수조직,채용면역형광법검측CXCL13화Neun적공표체정황이급성형효질세포활화정황.채용Western blot법측정CXCL13화GFAP적단백표체,채용RT-PCR법측정CXCL13화GFAP적mRNA표체.결과 여S조비교,NP、NS조화CS조술후각시점궤계통역하강,척수CXCL13화GFAP적단백급기mRNA표체상조(P＜0.05)；여NP조비교,CS조술후각시점궤계통역승고,척수CXCL13화GFAP적단백급기mRNA표체하조(P＜0.05),NS조각시점궤계통역、척수CXCL13화GFAP적단백급기mRNA표체차이무통계학의의(P＞0.05).결론 척수CXCL13통과격활성형효질세포삼여대서신경병이성통적형성화유지.
Objective To evaluate the role of chemokine CXC ligand 13 (CXCL13) in spinal cord in neuropathic pain (NP) in rats.Methods One hundred and eight male Sprague-Dawley rats,weighing 150-200 g,were randomly divided into 4 groups (n =27 each):sham operation group (group S),group NP,small interference RNA (siRNA) negative control group (group NS) and CXCL13-siRNA group (group CS).The animals were anesthetized with intraperitoneal ketamine 50 mg/kg.NP was induced by ligation of L5 spinal nerve (SNL) in groups NP,NS and CS.L5 spinal nerve was only exposed but not occluded in group S.CXCL13-siRNA lentivirus and siRNA negative control lentivirus were injected intrathecally in groups CS and NS,respectively.Mechanical pain threshold was measured at 3,7 and 14 days after SNL.Then the rats were sacrificed and L4-6 segments of the spinal cord were obtained for determination of coexpression of CXCL13 and Neun (by immunofluorescence),activation of astrocytes,and expression of CXCL13 and glial fibrillary acidic protein (GFAP) protein (by Western blot) and mRNA (by RT-PCR) in spinal cord tissues.Results Compared with group S,mechanical pain threshold was significantly decreased and the expression of CXCL13 and GFAP protein and mRNA was up-regulated at each time point after operation in groups NP,NS and CS (P ＜ 0.05).Compared with group NP,mechanical pain threshold was significantly increased and the expression of CXCL13 and GFAP protein and mRNA was down-regulated at each time point after operation in group CS (P ＜ 0.05).There was no significant difference in the indexes mentioned above at each time point after operation between groups NP and NS (P ＞ 0.05).Conclusion CXCL13 is involved in the development and maintenance of NP in rats via activation of astrocytes.