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甲状腺激素对氧化型低密度脂蛋白诱导的巨噬细胞功能紊乱的影响

宁瑜¹, 张铭^1,*, 杜芸辉², 张慧娜², 李林忆², 秦彦文², 温婉婉¹, 赵全明¹

¹首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所心内科，北京 100029；²首都医科大学附属北京安贞医院-北京市心肺血管疾病研究所上气道功能障碍相关心血管疾病研究室，北京 100029

摘要

甲状腺功能减退症(甲减)患者的动脉粥样硬化(atherosclerosis, AS)及冠心病风险指数显著增加，但机制不清楚。巨噬细胞功能紊乱是促进AS斑块形成及发展的重要环节，本研究旨在探讨甲状腺激素对巨噬细胞功能的直接影响，为甲减相关AS的机制研究提供新思路。用氧化型低密度脂蛋白(oxidized low-density lipoprotein, oxLDL)诱导小鼠单核巨噬细胞系RAW264.7，建立体外巨噬泡沫细胞模型，并观察不同浓度甲状腺素(thyroxine, T4)对巨噬泡沫细胞功能的改善效应。分别采用MTT法、划痕实验、β-半乳糖苷酶染色实验检测巨噬细胞增殖、迁移功能及细胞衰老情况；ELISA法检测巨噬细胞分泌肿瘤坏死因子-α (tumor necrosis factor-α, TNF-α)、单核细胞趋化蛋白-1 (monocyte chemoattractant protein-1, MCP-1)及白细胞介素-1β (interleukin-1β, IL-1β)情况；Western blot检测参与巨噬细胞增殖、迁移等过程的粘着斑激酶(focal adhesion kinase, FAK)的磷酸化水平。结果显示：oxLDL可显著抑制巨噬细胞增殖和迁移、促进其衰老及分泌 TNF-α、MCP-1和IL-1β，而T4可浓度依赖性逆转oxLDL对巨噬细胞上述功能的影响；oxLDL可使巨噬细胞磷酸化FAK蛋白表达上调，而T4可浓度依赖性降低FAK蛋白磷酸化水平。上述结果提示，T4可呈浓度依赖性地调控巨噬细胞增殖、迁移、衰老及炎性因子分泌等功能。

关键词： 甲状腺激素; 氧化型低密度脂蛋白; 巨噬细胞; 甲状腺功能减退症; 动脉粥样硬化

分类号：R363；R581.2；R541.4

Effects of thyroid hormone on macrophage dysfunction induced by oxidized low-density lipoprotein

NING Yu¹, ZHANG Ming^1,*, DU Yun-Hui², ZHANG Hui-Na², LI Lin-Yi², QIN Yan-Wen², WEN Wan-Wan¹, ZHAO Quan-Ming¹

¹Department of Cardiology, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing 100029, China；²Key Laboratory of Upper Airway Dysfunction-related Cardiovascular Diseases, Beijing Anzhen Hospital, Capital Medical University, Beijing Institute of Heart Lung and Blood Vessel Diseases, Beijing 100029, China

Abstract

It has been recognized that patients with hypothyroidism have higher risks of atherosclerosis and coronary heart disease, however, the mechanisms are largely unknown. Considering that macrophage dysfunction plays an important role in the formation and development of atherosclerosis plaques, this study aimed to investigate the direct effects of thyroid hormone on macrophage functions and to provide new insight for the mechanism of hypothyroid atherosclerosis. RAW264.7 cells (mouse leukaemic monocyte macrophage cell line) were incubated with oxidized low-density lipoprotein (oxLDL) to establish macrophage foam cells model in vitro, and the protective effects of different concentration of thyroxine (T4) on the macrophage foam cells function were explored. The proliferation, migration and cell aging of macrophages were detected by MTT method, scratch test and β-galactosidase staining respectively. The ELISA method was used to detect the secretion of tumor necrosis factor-α (TNF-α), monocyte chemoattractant protein-1 (MCP-1), and interleukin-1β (IL-1β). Western blot analysis was applied to measure the phosphorylation of focal adhesion kinase (FAK), which was required for the process of proliferation and migration of macrophages. The results showed that oxLDL significantly inhibited the macrophage proliferation and migration, induced cell senescence, and promoted the secretion of TNF-α, MCP-1, and IL-1β; while T4 reversed those effects of oxLDL on macrophage in a concentration-dependent manner. Moreover, oxLDL increased the phosphorylation of FAK in macrophage, while T4 concentration-dependently reversed the effect. These results suggest that T4 modulates macrophage proliferation, migration, senescence, and secretion of inflammation factors in a concentration-dependent way.

Key words: Thyroid hormone; oxidized low-density lipoprotein; Macrophage; hypothyroidism; atherosclerosis

收稿日期：2017-09-16　　录用日期：2018-01-30

通讯作者：张铭　　E-mail: mzcap@163.com

DOI: 10.13294/j.aps.2018.0015

引用本文：

宁瑜, 张铭, 杜芸辉, 张慧娜, 李林忆, 秦彦文, 温婉婉, 赵全明. 甲状腺激素对氧化型低密度脂蛋白诱导的巨噬细胞功能紊乱的影响[J]. 生理学报 2018; 70 (2): 141-148.

NING Yu, ZHANG Ming, DU Yun-Hui, ZHANG Hui-Na, LI Lin-Yi, QIN Yan-Wen, WEN Wan-Wan, ZHAO Quan-Ming. Effects of thyroid hormone on macrophage dysfunction induced by oxidized low-density lipoprotein. Acta Physiol Sin 2018; 70 (2): 141-148 (in Chinese with English abstract).