摘要
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14d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Westernblot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。
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 CXCLI3 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.
出处
《中华麻醉学杂志》
CAS
CSCD
北大核心
2013年第5期569-572,共4页
Chinese Journal of Anesthesiology
