文章摘要
杨少兵 ，胡柳 ，迟晓慧 ，张志发.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[J].神经损伤功能重建,2021,16(2):67-70
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
  
DOI：
中文关键词: 血神经屏障  神经病理性疼痛  Claudin-1  大鼠
英文关键词: blood nerve barrier  neuropathic pain  Claudin-1  rat
基金项目:湖北省自然科 学基金（No. 20 17CFB171）； 中央高校基本 科研业务费专 项资金（No. 20 17KFYXJJ128）
作者单位
杨少兵a ，胡柳b ，迟晓慧a ，张志发a 华中科技大学 同济医学院附 属同济医院 a. 麻醉学教研室， b.健康管理中 心 
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中文摘要:
      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 mRNA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。
英文摘要:
      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 1d 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 destruction of the BNB in the sciatic nerve.
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