神经病理性疼痛大鼠中脑导水管周围灰质腹外侧区NLRP3炎症小体激活水平研究  

Activation levels of NLRP3 inflammasomes in the ventrolateral periaqueductal gray of rats with neuropathic pain

作　　者：傅丹泓 彭梅静 黄海英 高贤伟 FU Dan-hong;PENG Mei-jing;HUANG Hai-ying;GAO Xian-wei(Department of Anesthesiology,Rehabilitation Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fuzhou Fujian 350003,China;Fujian Key Laboratory of Rehabilitation Technology,Fuzhou Fujian 350003,China)

机构地区：[1]福建中医药大学附属康复医院麻醉科,福建福州350003 [2]福建省康复技术重点实验室,福建福州350003

出　　处：《局解手术学杂志》2022年第2期108-113,共6页Journal of Regional Anatomy and Operative Surgery

基　　金：中央引导地方科技发展专项(2018L3009)。

摘　　要：目的探讨核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体在神经病理性疼痛大鼠中脑导水管周围灰质腹外侧区(vlPAG)的表达水平及其作用。方法选取SPF级SD大鼠20只,采用随机数字表法分为假手术组(Sham组)与慢性坐骨神经压迫性损伤组(CCI组),每组10只。于手术前后测定2组机械痛阈与热痛阈;Western blot检测2组NLRP3炎症小体各组分蛋白表达水平;RT-PCR检测2组NLRP3炎症小体各组分基因mRNA表达水平;酶联免疫吸附实验(ELISA)检测2组炎症因子白介素-lβ(IL^(-1)β)及白介素-18(IL^(-1)8)表达水平;免疫荧光检测2组中NLRP3炎症小体在vlPAG的分布情况,并与小胶质细胞标志物IBA1进行共定位分析。结果与Sham组相比,CCI组术后3 d、7 d、14 d、21 d机械痛阈与热痛阈均明显降低(P<0.001);与Sham组相比,CCI组核心蛋白NLRP3、效应蛋白Caspase-1 p10及前体蛋白pro-Caspase-1表达水平均明显升高(P<0.001);与Sham组相比,CCI组NLRP3(P<0.001)、Caspase-1(P<0.001)及IL^(-1)β(P<0.01)mRNA表达水平均明显升高,炎症因子IL^(-1)β(P<0.001)与IL^(-1)8(P<0.01)表达水平也明显升高;与Sham组相比,CCI组在vlPAG的NLRP3阳性细胞数增加,且与IBA1存在共表达。结论神经病理性疼痛大鼠vlPAG中NLRP3炎症小体被激活且表达水平增高,提示NLRP3炎症小体激活水平与神经病理性疼痛大鼠痛阈下降相关。Objective To detect the expression levels of nucleotide-binding oligomerization domain-like receptor protein 3(NLRP3)inflammasomes in the ventrolateral periaqueductal gray(vlPAG)of rats with neuropathic pain and its role.Methods A total of 20 SPF SD rats were selected and divided into the Sham operation group(the Sham group)and the chronic constriction injury of the sciatic nerve group(the CCI group)by random number table method,with 10 rats in each group.The mechanical pain threshold and thermal pain threshold in the two groups were measured before and after surgery.The protein expression levels of each component of the NLRP3 inflammasomes in the two groups were detected by Western blot.The mRNA expression levels of each component of the NLRP3 inflammasomes in the two groups were detected by RT-PCR.The expression levels of inflammatory factors interleukin-1β(IL^(-1)β)and interleukin-18(IL^(-1)β)in the two groups were detected by enzyme-linked immunosorbent assay(ELISA).The distribution of NLRP3 inflammasomes in vlPAG in the two groups was detected by immunofluorescence,and co-localization analysis was performed with the microglia marker IBA1.Results Compared with the Sham group,the mechanical pain threshold and thermal pain threshold in the CCI group were significantly decreased 3 days,7 days,14 days,and 21 days after surgery(P<0.001).Compared with the Sham group,the expression levels of core protein NLRP3,effector protein Caspase-1 p10 and precursor protein pro-Caspase-1 in the CCI group were significantly increased(P<0.001).Compared with the Sham group,the mRNA expression levels of NLRP3(P<0.001),Caspase-1(P<0.001)and IL^(-1)β(P<0.01)in the CCI group were significantly increased,as well as the expression levels of inflammatory factors IL^(-1)β(P<0.001)and IL^(-1)8(P<0.01).Compared with the Sham group,the number of NLRP3 positive cells in vlPAG was increased in the CCI group,and NLRP3 was co-expressed with IBA1.ConclusionNLRP3 inflammasomes in vlPAG of rats with neuropathic pain are activated and the expre

关 键 词：神经病理性疼痛 中脑导水管周围灰质腹外侧区 NLRP3炎症小体 炎症因子 慢性坐骨神经压迫性损伤 

分 类 号：R741.02[医药卫生—神经病学与精神病学]