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

MIN Jie; LI Hong

doi:10.16305/j.1007-1334.2022.2112099

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

更新时间：2022-12-22

- Regulatory effect of miR⁃155 on progress of the subacute thyroditis inflammatory response
- Shanghai Journal of Traditional Chinese Medicine Vol. 56, Issue 12, Pages: 27-33(2022)
- 作者机构：
  
  1.上海中医药大学附属龙华医院内分泌科（上海 200032）
- 作者简介：
- 基金信息：
- DOI：10.16305/j.1007-1334.2022.2112099
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闵婕,李红.miR⁃155在亚急性甲状腺炎炎症进展中的调控作用研究[J].上海中医药杂志,2022,56(12):27-33.

MIN Jie,LI Hong.Regulatory effect of miR⁃155 on progress of the subacute thyroditis inflammatory response[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(12):27-33.
闵婕,李红.miR⁃155在亚急性甲状腺炎炎症进展中的调控作用研究[J].上海中医药杂志,2022,56(12):27-33. DOI： 10.16305/j.1007-1334.2022.2112099.

MIN Jie,LI Hong.Regulatory effect of miR⁃155 on progress of the subacute thyroditis inflammatory response[J].Shanghai Journal of Traditional Chinese Medicine,2022,56(12):27-33. DOI： 10.16305/j.1007-1334.2022.2112099.

摘要

目的,2,研究miR⁃155对亚急性甲状腺炎（SAT）炎症进展的调控作用。,方法,2,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、25 μg·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,-,1β ,mRNA表达水平，ELISA法检测IL-6、IL-8、IL-1β释放水平。,结果,2,①SAT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,-,1β ,mRNA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,2,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。

Abstract

Objective,2,To investigate the regulatory effect of miR-155 on the progress of the subacute thyroditis （SAT） inflammatory response.,Methods,2,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-α with different concentrations （0， 1， 5， 25 μg·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,-,1β, mRNA and miR-155， and ELISA was performed to detect protein levels of IL-6， IL-8， and IL-1β.,Results,2,①The expression levels of miR-155,，, IL-6 and TNF-α in 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-α could upregulate the expression level of miR-155， and induce the expression and release of ,IL,-,6,， ,IL-8, and ,IL,-,1β, mRNA， 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-1β showed a marked increase after overexpression of miR-155 （,P,<,0.05）； On the other hand， TNF-α induced mRNA expression and release of IL-6， IL-8 and IL-1β were suppressed after suppression of miR-155 （,P,<,0.05）.,Conclusions,2,miR-155,plays a role in promoting inflammation in the SAT， which provides a new direction for the target research of traditional Chinese medicine in treating SAT.

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

Keywords

subacute thyroiditismiR-155miRNANthy-ori 3-1 cellinflammation

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