miR-155在亚急性甲状腺炎炎症进展中的调控作用研究  

Regulatory effect of miR-155 on progress of the subacute thyroditis inflammatory response

作　　者：闵婕 李红[1] MIN Jie;LI Hong(Department of Endocrinology,Long Hua Hospital,Shanghai University of Traditional Chinese Medicine,Shanghai 200032,China)

机构地区：[1]上海中医药大学附属龙华医院内分泌科,上海200032

出　　处：《上海中医药杂志》2022年第12期27-33,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81373617);上海市卫健委中医药科研项目(2020 JQ001);上海市科委“科技创新行动计划”项目(19140904600);上海市科委2018年度医学引导类(中、西医)科技支撑项目(18401900800)。

摘　　要：目的研究miR⁃155对亚急性甲状腺炎(SAT)炎症进展的调控作用。方法收集SAT患者39例、同期健康体检者36例,分别设为SAT组、正常组,检测研究对象血清miR⁃155表达水平及细胞因子[白介素-2(IL-2)、白介素-6(IL-6)、白介素-10(IL-10)、肿瘤坏死因子-α(TNF-α)]含量,并对miR⁃155表达水平与炎症(白细胞计数、红细胞沉降率、超敏C反应蛋白)、甲状腺功能[甲状腺球蛋白(TG)、游离三碘甲腺原氨酸(FT_(3))、游离甲状腺素(FT_(4))、促甲状腺激素(TSH)]指标进行相关性分析。TNF-α(0、1、5、25μg·L^(-1))诱导人甲状腺正常细胞(Nthy-ori 3-1),构建甲状腺细胞炎症模型,利用脂质体转染oligoRNA调控细胞内miR⁃155表达水平,荧光显微镜鉴定转染效率,PCR法检测miR⁃155及IL-6、IL-8、IL-1βmRNA表达水平,ELISA法检测IL-6、IL-8、IL-1β释放水平。结果①SAT患者血清中miR⁃155水平及IL-6、TNF-α含量高于正常人群(P<0.05)。②SAT患者血清中miR⁃155的表达水平与IL-6(P<0.01,r=0.859)、TNF-α(P<0.01,r=0.958)、FT_(3)(P<0.05,r=0.641)、FT_(4)(P<0.01,r=0.775)呈正相关。③TNF-α可以上调Nthy-ori 3-1的miR⁃155表达水平,诱导IL-6、IL-8、IL-1βmRNA表达及释放,并存在浓度依赖趋势。④转染oligoRNA 6 h后,转染效率达到约90%;与阴性对照组相比,miR⁃155模拟物(mimics)(30 nmol·L^(-1))可以显著上调miR⁃155(P<0.05),miR⁃155抑制物(inhibitor)(100 nmol·L^(-1))可以显著下调miR⁃155(P<0.05)。⑤过表达miR⁃155后,IL-1β、IL-6及IL-8的mRNA表达及释放升高(P<0.05);下调miR⁃155表达后,TNF-α诱导的IL-1β、IL-6及IL-8的mRNA表达及释放被抑制(P<0.05)。结论miR⁃155在SAT中有促进炎症进展的作用,为中医药治疗SAT作用靶点研究提供了新的方向。Objective To investigate the regulatory effect of miR-155 on the progress of the subacute thyroditis(SAT)inflammatory response.Methods We collected 39 cases of subacute thyroiditis and 36 cases of normal control,detected circulating miR-155 expression level and the protein levels of inflammatory cytokines(IL-2,IL-6,IL-10,TNF-α)of subjects.Correlation analysis was carried out on the circulating miR-155 expression level of SAT patients with related inflammation indicators(WBC,ESR,CRP)and thyroid function indicators(TG,FT_(3),FT_(4),TSH).Nthy-ori 3-1 cells were induced by TNF-αwith different concentrations(0,1,5,25μg·L^(-1))to establish a thyroid cell inflammation model,and the expression level of miR-155 was regulated by oligoRNA,which was transfected by lipofectamine 2000.Fluorescence microscope was used to identify transfection efficiency,qRT-PCR was performed to detect the expression of IL-6,IL-8,IL-1βmRNA and miR-155,and ELISA was performed to detect protein levels of IL-6,IL-8,and IL-1β.Results①The expression levels of miR-155,IL-6 and TNF-αin SAT patients were significantly higher than those of healthy control participants(P<0.05).②The expression levels of miR-155 in serum were positively correlated with IL-6(P<0.01,r=0.859),TNF-α(P<0.01,r=0.958),FT_(3)(P<0.05,r=0.641)and FT_(4)(P<0.01,r=0.775)in SAT patients.③In Nthy-ori 3-1 cells,TNF-αcould upregulate the expression level of miR-155,and induce the expression and release of IL-6,IL-8 and IL-1βmRNA,which had a concentration-dependent trend.④The transfection efficiency reached about 90%after 6 hours of oligoRNA transfection.Compared with the condition in the negative control,miR-155 mimics(30 nmol·L^(-1))could significantly upregulate the expression level of miR-155(P<0.05),while miR-155 inhibitor(100 nmol·L^(-1))could significantly downregulate the expression level of miR-155(P<0.05).⑤mRNA expression and release of IL-6,IL-8 and IL-1βshowed a marked increase after overexpression of miR-155(P<0.05);On the other hand,TNF-αinduced mRNA ex

关 键 词：亚急性甲状腺炎 miR⁃155 微小核糖核酸 人甲状腺正常细胞 炎症 

分 类 号：R581.4[医药卫生—内分泌]