小胶质细胞在Tourette综合征中活化及作用机制研究  

Activation and mechanism of microglia in Tourette syndrome

作　　者：张小玲 刘秀梅 ZHANG Xiaoling;LIU Xiumei(Department of Pediatrics,The 900th Hospital of Joint Logistic Support Force,Fuzhou,Fujian 350025,China;Department of Developmental and Behavioral Pediatrics,Fujian Children′s Hospital)

机构地区：[1]解放军联勤保障部队第900医院儿科,福建福州350025 [2]福建省儿童医院发育行为科

出　　处：《中国儿童保健杂志》2024年第12期1316-1321,共6页Chinese Journal of Child Health Care

基　　金：福建省自然科学基金面上项目(2021J01424)。

摘　　要：目的探讨小胶质细胞(MG)在3,3′-亚氨基二丙腈(IDPN)诱导的Tourette综合征(TS)大鼠中的活化及作用机制。方法雄性Sprague-Dawley大鼠36只,随机分为TS组和假用药组(Sham组),每组18只,TS组腹腔注射IDPN[300mg/(kg·d),7d],Sham组腹腔注射生理盐水[5mL/(kg·d),7d]。使用免疫荧光双染检测大鼠纹状体组织M1型MG标记蛋白诱导型一氧化氮合酶(iNOS)表达及与MG活化特异性标记物离子钙接头蛋白-1(Iba-1)共定位的情况,M2型MG标记蛋白精氨酸酶-1(Arg-1)表达及与Iba-1共定位的情况;使用酶联免疫吸附法(ELISA)、实时荧光定量PCR(qRT-PCR)对大鼠纹状体组织中iNOS、Arg-1的表达水平进行定量,同时检测炎症因子肿瘤坏死因子α(TNF-α)、白细胞介素-6(IL-6)、白细胞介素-10(IL-10)的表达水平。结果大鼠纹状体组织免疫荧光双染显示,相比Sham组,TS组大鼠纹状体组织中Iba-1、iNOS的阳性表达均有所增强且呈现共定位,Arg-1的阳性表达有所降低且与Iba-1呈现共定位。ELISA结果显示,相比Sham组,TS组M1型MG标记蛋白iNOS含量显著升高(t=5.796,P<0.001),M2型MG标记蛋白Arg-1含量显著降低(t=4.348,P<0.01),炎症因子TNF-α、IL-6、IL-10含量均显著升高(t=5.654、5.748、8.231,P<0.001);qRT-PCR结果显示,与Sham组相比,TS组M1型MG标记蛋白iNOS的mRNA表达水平显著升高(t=9.914,P<0.001),M2型MG标记蛋白Arg-1的mRNA表达水平显著降低(t=4.390,P<0.01),炎症因子TNF-α、IL-6、IL-10的mRNA表达水平均显著升高(t=12.056、14.147、13.350,P<0.001)。结论IDPN诱导的TS大鼠存在MG活化,且M1型MG上调,M2型MG下调,介导神经炎症。Objective To investigate the activation and mechanism of microglia(MG)in Tourette syndrome(TS)rats induced by 3,3′-aminodipropionitrile(IDPN).Methods Thirty six male Sprague Dawley rats were randomly divided into a TS group and a Sham group(Sham group),with 18rats in each group.The TS group received intraperitoneal injection of IDPN for 7days,with the dose of 300mg/(kg·d),while the Sham group received intraperitoneal injection of physiological saline for 7days,with the dose of 5mL/(kg·d).Double immunofluorescence staining was used to detected the expression of inducible nitric oxide synthase(iNOS),a marker protein of M1type microglia,and its co-localization with Iba-1(a specific marker for microglial activation)in rat striatum,as well as the expression of arginase 1(Arg-1)and its co-location with Iba-1 in M2 microglia.The expression levels of iNOS,Arg-1 in rat striatal tissues were quantified by enzyme-linked immunosorbent measurement(ELISA),real-time PCR(qRT-PCR),meanwhile the expression levels of inflammatory cytokines tumor necrosis factorα(TNF-α),interleukin-6(IL-6),and interleukin-10(IL-10)were detected.Results Immunofluorescence double staining showed that the positive expressions of Iba-1and iNOS in striatum of TS group were enhanced and co-located compared with Sham group,while the positive expression of Iba-1and Arg-1in striatum of TS group decreased and showed co-localization.The ELISA results showed that compared with the Sham group,the TS group had a significant increase in the content of M1type MG marker protein iNOS(t=5.796,P<0.001),a significant decrease in the content of M2type MG marker protein Arg-1(t=4.348,P<0.01),and a significant increase in inflammatory factors,including TNF-α,IL-6and IL-10(t=5.654,5.748,8.231,P<0.001).The qRT-PCR results showed that compared with the Sham group,the mRNA expression level of iNOS,a M1type MG marker protein,was significantly increased in the TS group(t=9.914,P<0.001),while the mRNA expression level of Arg-1,a M2type MG marker protein,was significantly r

关 键 词：TOURETTE综合征 小胶质细胞 神经免疫 炎症因子 

分 类 号：R749.94[医药卫生—神经病学与精神病学]