菖蒲郁金汤对抽动-秽语综合征模型大鼠小胶质细胞的异常活化和神经炎症的影响  

Effects of Changpu Yujin Tang(菖蒲郁金汤)on abnormal activation of microglia and neuroinflammation in a rat model of Tourette syndrome

作　　者：罗文珍 尚菁[1] 史正刚[1] 冯鹏 孙治前 高蕾 王梓安 LUO Wenzhen;SHANG Jing;SHI Zhenggang;FENG Peng;SUN Zhiqian;GAO Lei;WANG Zi’an(Clinical College of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou,Gansu,730101,China)

机构地区：[1]甘肃中医药大学中医临床医学院,甘肃兰州730101

出　　处：《中医儿科杂志》2023年第6期1-9,共9页Journal of Pediatrics of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81960886);甘肃省高等学校创新基金项目(2020A-081);甘肃省自然科学基金项目(20JR10RA324)。

摘　　要：目的探讨菖蒲郁金汤通过调节非编码小分子RNA Let7i(microRNA Let7i,Let7i)对3,3′-亚氨基二丙腈(3,3′-Lminodipropionitrile,IDPN)诱导的抽动-秽语综合征(TS)模型大鼠中小胶质细胞(microglia,MG)的异常活化及神经炎症的影响。方法将40只SPF级Sprague-Dawley 3周龄雄性大鼠随机分为空白组、模型组、硫必利组和菖蒲郁金汤组,每组10只。腹腔注射IDPN制备TS模型,硫必利组给予硫必利混悬液,菖蒲郁金汤组给予菖蒲郁金汤药液,空白组和模型组给予等体积生理盐水,按15 mL/(kg·d)灌胃。干预28 d后,采用免疫荧光法检测MG活化情况,FISH检测Let7i的表达,免疫组织化学法(IHC)、WB法和Rt-PCR法检测纹状体Toll样受体4(Toll-Like receptor4,TLR4)、髓样分化初次应答因子88(myeloid differentiation factor 88,MYD88)、转录因子核因子κB(nuclear factor-κB,NF-κB)蛋白和mRNA的表达。结果干预前,空白组大鼠纹状体中MG多呈静息状态,氧化还原性异丁醛(Iba1)受体表达很少,荧光强度较弱;模型组、硫必利组和菖蒲郁金汤组纹状体活化MG数量较空白组明显增加,荧光强度增强,差异有统计学意义(P<0.05)。干预后,与空白组比较,模型组Iba1表达仍较高,差异有统计学意义(P<0.05);与模型组比较,菖蒲郁金汤组Iba1表达减少,荧光面积减少,差异有统计学意义(P<0.05)。干预前,与空白组比较,模型组、硫必利组和菖蒲郁金汤组大鼠纹状体Let7i表达减少,差异有统计学意义(P<0.05);干预后,与模型组比较,硫必利组和菖蒲郁金汤组Let7i表达升高,差异有统计学意义(P<0.05),但2组间比较,差异无统计学意义(P>0.05)。干预前,模型组、硫必利组和菖蒲郁金汤组TLR4、MYD88、NF-κB表达及其蛋白水平较高,与空白组比较,差异有统计学意义(P<0.05)。干预后,模型组TLR4、MYD88、NF-κB表达及其蛋白水平较高,与空白组比较,差异有统计学意义(P<0.05);与模型组比较,硫必利组和菖蒲郁金�Objective To investigate the effect of Changpu Yujin Tang(菖蒲郁金汤)on the abnormal activa⁃tion of microglia(MG)and neuroinflammation in a rat model of Tourette syndrome(TS)induced by 3,3'⁃imino⁃dipropionic nitrile(IDPN)by regulating non coding small molecule RNA Let7i(Let7i).Methods Forty Sprague Dawley three⁃week⁃old male rats of SPF grade were randomly divided into a blank group,a model group,a Tiapride group,and a Changpu Yujin Tang(CYT)group,with 10 rats in each group.The TS model was prepared by intraper⁃itoneal injection of IDPN.The Tiapride group was given Tiapride suspension,the CYT group was given Changpu Yu⁃jin Tang,and the blank group and model group were given equal volume of physiological saline at a rate of 15 mL/(kg·d)by gavage.After 28 days of intervention,MG activation was detected using immunofluorescence as⁃say,expression of Let7i was detected using FISH,and Toll like receptor 4(TLR4),myeloid differentiation first re⁃sponse factor 88(MYD88),and transcription factor nuclear factor were detected using IHC,WB,and Rt PCR meth⁃ods⁃κB(NF⁃κB)expression of proteins and mRNA.Results Before the intervention,MG in the striatum of the blank group rats was mostly in a resting state,with little expression of Iba1 receptor and weak fluorescence intensity.The number of activated MG in the striatum of the model group,the Thiopride group,and the CYT group was signif⁃icantly increased compared to the blank group,and the fluorescence intensity was enhanced,with a statistically sig⁃nificant difference(P<0.05).After intervention,compared with the blank group,the model group still had a higher expression of Iba1 in microglia,with a statistically significant difference(P<0.05).Compared with the model group,the number of Iba1 microglia and fluorescence area in the CYT group decreased,and the difference was statistically significant(P<0.05).Before intervention,compared with the blank group,the expression of Let7i in the striatum of rats in the TS model group,Tiapride group,and CYT g

关 键 词：菖蒲郁金汤 抽动-秽语综合征 非编码小分子RNA Let7i 小胶质细胞 神经炎症 实验研究 

分 类 号：R749.94[医药卫生—神经病学与精神病学]