甲状腺激素对巨噬细胞调控作用的实验研究  

Study on the Regulatory Effect of Thyroxine on Macrophages

作　　者：陈子维 蔡东成 聂宇[1] CHEN Zi-wei;CAI Dong-cheng;NIE Yu(State Key Laboratory of Cardiovascular Disease,Funwai Hospital,Chinese Academy of Medical Sciences,Peking Union Medical College,Bejing 100037,China)

机构地区：[1]中国医学科学院北京协和医学院阜外医院心血管疾病国家重点实验室,北京市100037

出　　处：《中国分子心脏病学杂志》2021年第5期4236-4241,共6页Molecular Cardiology of China

基　　金：国家自然科学基金(81770308)。

摘　　要：目的探讨甲状腺激素对巨噬细胞毒性、泡沫化、极化和迁移功能的调控作用。方法采用CCK-8比色法测定RAW 264.7小鼠巨噬细胞活性;采用流式细胞术检测甲状腺激素对巨噬细胞凋亡的影响;使用氧化低密度脂蛋白(ox-LDL)诱导巨噬细胞泡沫化,用油红O染色法检测甲状腺激素对巨噬细胞泡沫化的影响;使用实时荧光定量PCR(RT-qPCR)检测甲状腺激素对RAW 264.7小鼠巨噬细胞M1型和M2型极化的影响;采用划痕实验检测甲状腺激素对小鼠骨髓来源的巨噬细胞(BMDM)迁移能力的影响。结果CCK-8比色法和流式细胞术结果提示,与对照组相比,甲状腺激素在基础状态下可以减少巨噬细胞凋亡和坏死,增强巨噬细胞的活性;油红O染色结果提示,与对照组相比,甲状腺激素可以促进巨噬细胞泡沫化,且具有剂量依赖性,甲状腺激素浓度越高,巨噬细胞泡沫化程度越高;RT-qPCR结果提示,与对照组相比,甲状腺激素可以促进巨噬细胞向M1型极化,抑制巨噬细胞向M2型极化;与对照组相比,甲状腺激素能够促进脂多糖(LPS)和γ干扰素(IFN-γ)诱导的M1型巨噬细胞炎症因子肿瘤坏死因子-α(TNF-α)(P<0.01)和M1型极化标志物诱导型一氧化氮合酶(iNOS)、细胞色素C氧化酶亚基Ⅱ(COX-2)的表达(P<0.05),同时抑制白细胞介素-4(IL-4)诱导的M2型巨噬细胞标志物CD206的表达(P<0.05);划痕实验结果显示,甲状腺激素可以促进巨噬细胞迁移。结论甲状腺激素可以促进巨噬细胞向M1型极化,抑制其向M2型极化。而且甲状腺激素可以减少基础状态下巨噬细胞的凋亡和坏死,能够增强巨噬细胞的活性,增强巨噬细胞泡沫化,促进巨噬细胞迁移。Objective To investigate the regulatory effect of thyroxine on macrophage cytotoxicity,apoptosis,foaming,polarization and migration.Methods CCK-8 colorimetry assays was used to determine cell viability of RAW 264.7 cells.Flow cytometry was used to detect cell apoptosis.Oxidized low density lipoprotein(ox-LDL)was used to induce macrophage foaming,and the intracellular lipid profile was observed by oil red O staining.The effect of thyroxine on M1 and M2 polarization were measured by real-time fluorescent quantitative PCR(RT-qPCR).Cell scratch assay was used to detect macrophage migration.Results Thyroxine reduced macrophage apoptosis and necrosis in basal state,and compared with the control group,thyroxine enhanced cell viability.The intracellular lipid was increased in thyroxine treatment group,and the degree of cell foaming was increased as the concentration of thyroxine increases.Thyroxine increased the expression of tumor necrosis factor-α(TNF-α)(P<0.01),M1 marker including inducible nitric oxide synthase(iNOS)and cytochrome C oxidase subunitⅡ(COX-2)induced by lipopolysaccharide(LPS)and interferonγ(IFN-γ),but reduced the expression of M2 marker CD206 induced by interleukin-4(IL-4)(P<0.05).Thyroxine could promote macrophage migration.Conclusions Thyroxine promote the polarization of macrophages to M1-type and inhibit M2 polarization,and reduce macrophage apoptosis and necrosis in basal state,and could enhance cell viability,enhance the foaming of macrophages,promote macrophage migration.

关 键 词：甲状腺激素 巨噬细胞 细胞毒性 泡沫化 极化 

分 类 号：R392[医药卫生—免疫学]