血神经屏障和Claudin-1蛋白在大鼠慢性神经病理性疼痛中的作用  

Effect of Blood Nerve Barrier and Claudin-1 on Chronic Neuropathic Pain in Rats

作　　者：杨少兵[1] 胡柳[2] 迟晓慧[1] 张志发[1] YANG Shao-bing;HU Liu;CHI Xiao-hui;ZHANG Zhi-fa(Department of Anesthesiology and Pain Medicine,Tongji Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China;Physical Examination Center,Tongji Hospital of Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China)

机构地区：[1]华中科技大学同济医学院附属同济医院麻醉学教研室,武汉430030 [2]华中科技大学同济医学院附属同济医院健康管理中心,武汉430030

出　　处：《神经损伤与功能重建》2021年第2期67-70,共4页Neural Injury and Functional Reconstruction

基　　金：湖北省自然科学基金(No.2017CFB171);中央高校基本科研业务费专项资金(No.2017KFYXJJ128)。

摘　　要：目的:观察慢性神经病理性疼痛大鼠坐骨神经的血神经屏障(BNB)变化,并探讨紧密连接蛋白Claudin-1在其中的作用。方法:健康雌性SD大鼠分为3组:对照组(Control组)、假手术组(Sham组)和慢性坐骨神经压迫损伤组(CCI组)。对照组不做处理;Sham组仅分离暴露大鼠右侧坐骨神经,不结扎;CCI组暴露大鼠右侧坐骨神经并用丝线结扎。术后1 d,采用伊文斯蓝蛋白染剂(EBA)观察大鼠坐骨神经BNB通透性的变化;免疫荧光染色法观察坐骨神经内的Claudin-1蛋白分布。术前及术后6 h、1 d、7 d和14 d测定各组大鼠后肢机械痛阈值,RT-PCR法测定Claudin-1 m RNA转录水平,Western blot法测定Claudin-1蛋白表达水平。结果:与其他2组相比,CCI组大鼠术后1 d右侧后肢机械痛阈值降低,术后7 d降至最低,并且一直持续到术后14 d(均P<0.05);术后1 d,CCI组EBA含量显著增高(P<0.05);术后6 h、1 d、7 d和14 d CCI组大鼠坐骨神经内Claudin-1 mRNA转录水平和Claudin-1蛋白表达水平均显著低于其他2组(均P<0.05);坐骨神经免疫荧光染色显示,Claudin-1蛋白主要分布于神经内膜和神经束膜,术后1 d,CCI组坐骨神经束膜中Claudin-1蛋白荧光强度明显低于其他2组(均P<0.05)。结论:慢性神经病理性疼痛大鼠坐骨神经BNB被破坏,通透性增加,神经内紧密连接蛋白Claudin-1蛋白表达水平减少。Objective:This study observed the changes in the blood nerve barrier(BNB)in rat chronic neuropathic pain and investigated the role of tight junction protein Claudin-1 in regulating it’s permeability.Methods:Healthy female SD rats were randomly divided into the control group,sham operation group(sham group),and chronic constriction injury of sciatic nerve group(CCI group).No treatment was given in the control group;the right sciatic nerves of rats were exposed but not ligated in the sham group;the sciatic nerves were separated and treated with four loose knots in the CCI group.The change in sciatic nerve permeability was observed by studying Evans blue-labelled albumin(EBA).The distribution of Claudin-1 in the sciatic nerve was detected by immunofluorescence staining.The right hindlimb mechanical pain threshold was measured at 6 h,1 d,7 d,and 14 d after operation.The transcription levels of Claudin-1 mRNA were detected by RT-PCR,and the expression levels of Claudin-1 protein were determined by Western blot.Results:Compared with the control group and sham group,the CCI group showed significantly decreased hindlimb mechanical pain thresholds beginning 1 d after operation,reaching its lowest point at 7 d,and maintaining this level until 14 d(all P<0.05).The CCI group showed significantly increased EBA penetration 1 d after operation(P<0.05).Compared with the other two groups,the CCI group showed a significant decrease in Claudin-1 mRNA transcription levels and Claudin-1 protein expression levels in the sciatic nerve at 6 h,1 d,7 d,and 14 d after operation(all P<0.05).Immunofluorescence staining of the sciatic nerve showed that Claudin-1 was mainly distributed within the endoneurium and perineurium.Compared with the other two groups,the CCI group displayed significantly reduced intensity in Claudin-1 protein fluorescence within the sciatic nerve perineurium 1 d after operation(both P<0.05).Conclusion:In rats with chronic neuropathic pain,the expression level of Claudin-1 protein is reduced,resulting in the destructi

关 键 词：血神经屏障 神经病理性疼痛 CLAUDIN-1 大鼠 

分 类 号：R741[医药卫生—神经病学与精神病学] R741.02[医药卫生—临床医学]