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硫化氢通过 Nrf 2/HO-1 对神经病理性疼痛的调控作用
田礼华,韩焕芝,张杨,谢克亮,陈红光
0
天津市武清区中医院疼痛科(天津 301700);山东德州市人民医院麻醉科(德州 253014);天津医科大学总医院麻醉科(天津 300054)
摘要:
目的:探讨 Nrf2/HO-1 在硫化氢减轻神经病理性疼痛中的调控作用。方法:选择雄性 SD 大鼠 64 只,采用随机数字表法随机分为对照组(Con 组)、神经病理性疼痛组(SNL 组)、神经病理性疼痛 +NaHS 组(SNL+NaHS 组)和神经病理性疼痛 +NaHS+Nrf2 siRNA 组(SNL+siRNA+NaHS 组)4 组。神经病理性疼痛模型采用脊神经结扎法(SNL)完成。Con 组、SNL 组和 SNL+NaHS 组接受处理后 7 d,收集各组大鼠脊髓膨大处,部分组织进行核蛋白和 RNA 的提取进行 Nrf2 的检测,部分组织进行免疫荧光 IBA1 和 Nrf2 的共定位检测。4 组大鼠在给予处理后的 1、2、3、5 和 7 d 进行行为学的检测,结束后收集大鼠脊髓膨大处进行 IBA1、Nrf2 和 HO-1 蛋白的检测,部分组织匀浆进行炎症因子 TNF-α和IL-1β 的检测。结果:与 Con 组比较,SNL 大鼠机械痛敏减少,热痛敏缩短,Nrf2 核蛋白和 mRNA 表达增加,免疫荧光结果 Nrf2 和 IBA1 蛋白表达及共定位表达增加,总蛋白 IBA1、Nrf2 和 HO-1 蛋白表达增加,TNF-α 和 IL-1β 表达增加; 与 SNL 组相比,SNL+NaHS 组机械痛敏增加,热痛敏延长,Nrf2 核蛋白和 mRNA 表达进一步增加,免疫荧光结果示 Nrf2 和 IBA1 蛋白表达及共定位表达进一步增加,总蛋白 IBA 蛋白表达减少,Nrf2 和 HO-1 蛋白表达进一步增加,TNF-α和IL-1β 表达减少;与 SNL+NaHS 组比较,SNL+siRNA+NaHS 组机械痛敏减少,热痛敏缩短,总蛋白 IBA1 蛋白表达增加、Nrf2 和 HO-1 蛋白表达减少,TNF-α和 IL-1β 表达增加。结论:NaHS 对 SNL 导致的神经病理性疼痛小胶质细胞的激活, 是通过调节 Nrf2/HO-1 信号通路实现的。
关键词:  神经病理性疼痛  小胶质细胞  硫化氢
DOI:10.3969/j.issn.1007-6948.2019.02.001
基金项目:
Regulation of Neuropathic Pain by H2S via Nrf2/HO-1 Pathway
TIAN Li-hua,HAN Huan-zhi,ZHANG Yang,XIE Ke-liang,CHEN Hong-guang
Department of Pain, Wuqing Hospital of Ttraditional Chinese Medicine of Tianjin City, Tianjin (301700), China
Abstract:
Objective To investigate the regulated role of Nrf2 /HO- 1 on H 2 S alleviating the microglia activation in neuropathic pain in rats. Methods Sixty-four adult male Sprague-Dawley rats were randomly divided into four groups: control group (group Con, n = 16 ), neuropathic pain group (group SNL, n = 16 ), neuropathic pain+NaHS group (group SNL+NaHS, n = 16 ) and neuropathic pain+NaHS+Nrf 2 siRNA group (SNL+siRNA+NaHS, n=16). The animal model of neuropathic pain was performed by spinal nerve ligation (SNL). The L5-6 spine cords were collected for the detection of Nrf2 protein and Nrf2 mRNA by Western blotting and RT-PCR and IBA1 and Nrf2 co-location by immuno?uorescence after 7 d of administration in group Con, group SNL and group SNL+NaHS. After 1 , 2 , 3 , 5 and 7 d of operation, MWT and TWL were measured, and after detection of behavioristics, the L5-6 spine cords were collected for the detection of IBA1, Nrf2 and HO-1 protein by Western blotting and cytokines TNF-α and IL-1β by ELISA. Results Compared with group Con, MWT and TWL were reduced, Nrf2 nucleoprotein and mRNA, co-location of Nrf2 and IBA1 expression, total protein IBA1, Nrf2 and HO-1, and cytokines TNF-α and IL-1β were increased in group SNL; compared with group SNL, MWT and TWL were improved in group SNL+NaHS; compared with group SNL+NaHS, MWT and TWL were reduced, Nrf2 nucleoprotein and mRNA, the co-location of Nrf2 and IBA1 expression, total protein Nrf 2 and HO- 1 , were reduced, and total protein IBA1, cytokines TNF-α and IL-1β were increased in group SNL+NaHS+siRNA. Conclusion NaHS could alleviate the microglia activation by Nrf2/HO-1 pathway in neuropathic pain.
Key words:  Neuropathic pain  microglia  H2S

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