健康人来源高密度脂蛋白对小鼠骨髓来源巨噬细胞炎症反应的调控作用研究

The regulatory effect of healthy human-derived high-density lipoprotein on inflammation in mouse bone marrow-derived macrophages

目的探讨健康人来源的高密度脂蛋白(HDL)对小鼠骨髓来源巨噬细胞炎症反应的调控作用。方法提取健康人的HDL,提取小鼠骨髓来源的巨噬细胞,观察HDL对脂多糖和白细胞介素4(IL-4)分别刺激的巨噬细胞炎症因子分泌的影响。提取小鼠主动脉平滑肌细胞,利用星形孢菌素将平滑肌细胞诱导为凋亡细胞碎片,观察HDL对平滑肌细胞碎片刺激的巨噬细胞炎症因子分泌的影响。根据巨噬细胞的刺激方式进行不同的实验分组,在脂多糖刺激的条件下,巨噬细胞分为对照组、HDL组、脂多糖组和脂多糖+HDL组;在IL-4的刺激的条件下,巨噬细胞分为对照组、IL-4组和IL-4+HDL组;在细胞碎片刺激的条件下,巨噬细胞分为对照组、碎片组和碎片+HDL组。结果脂多糖组巨噬细胞炎症因子肿瘤坏死因子(TNF)、γ干扰素、单核细胞趋化蛋白1(MCP-1)、IL-10、IL-6表达水平均高于对照组;脂多糖+HDL组TNF、γ干扰素、MCP-1、IL-10、IL-6表达水平均低于脂多糖组(均P<0.05)。IL-4组MCP-1表达水平高于对照组[(147±12)ng/L比(21±4)ng/L],IL-4+HDL组MCP-1表达水平[(112±11)ng/L]低于IL-4组,TNF表达水平高于IL-4组[(159.4±36.0)ng/L比(0.9±0.7)ng/L](均P<0.05)。碎片组TNF、γ干扰素、MCP-1、IL-10表达水平高于对照组;碎片+HDL组γ干扰素和IL-10表达水平低于碎片组,TNF和IL-6表达水平高于碎片组(均P<0.05)。结论HDL对巨噬细胞炎症因子的调控受外围环境的影响。在脂多糖刺激的环境下,HDL能够抑制巨噬细胞的炎症反应;在IL-4或者平滑肌碎片刺激的环境下,HDL对不同的炎症因子有着不同的调控作用。HDL作用复杂,在某些环境刺激下,可能同时存在促炎和抑炎的作用。

Objective To investigate the regulatory effect of healthy human-derived high-density lipoprotein(HDL)on the inflammatory of mouse bone marrow-derived macrophages.Methods The healthy human-derived HDL and mouse bone marrow-derived macrophages were extracted to observe the effect of HDL on the secretion of inflammatory factors in macrophages stimulated with lipopolysaccharide(LPS)or interleukin-4(IL-4)respectively.Smooth muscle cells were isolated from mouse aorta and induced them into apoptosis cell debris with staurosporine.The effect of HDL on the secretion of inflammatory factors in macrophages stimulated with smooth muscle cell debris was observed.According to the stimulation on macrophages,different experimental groups were conducted.Under the stimulation with LPS,macrophages were divided into control group,HDL group,LPS group and LPS+HDL group;under the stimulation with IL-4,macrophages were divided into control group,IL-4 group and IL-4+HDL group;under the stimulation with cell debris,macrophages were divided into control group,debris group and debris+HDL group.Results The expression levels of macrophage inflammatory factors tumor necrosis factor(TNF),interferon-γ,monocyte chemoattractant protein 1(MCP-1),IL-10,and IL-6 in the LPS group were higher than those in the control group;the expression levels of TNF,interferon-γ,MCP-1,IL-10 and IL-6 in the LPS+HDL group were lower than those in the LPS group(all P<0.05).The expression level of MCP-1 in the IL-4 group was higher than that in the control group[(147±12)ng/L vs(21±4)ng/L],and the expression level of MCP-1 in the IL-4+HDL group[(112±11)ng/L]was lower than that in the IL-4 group,and the expression level of TNF in the IL-4+HDL was higher than that in the IL-4 group[(159.4±36.0)ng/L vs(0.9±0.7)ng/L](all P<0.05).The expression levels of TNF,interferon-γ,MCP-1,and IL-10 in the debris group were higher than those in the control group;the expression levels of interferon-γand IL-10 in the debris+HDL group were lower than those in the debris group,and the expression levels of TNF and IL-6 in the debris+HDL group were higher than those in the debris group(all P<0.05).Conclusions The regulation of HDL in macrophage inflammatory factors is affected by the environment.In the stimulation with LPS,HDL inhibits the inflammatory response of macrophages.In the stimulation of IL-4 or smooth muscle cell debris,HDL has different regulatory effects on different inflammatory factors.The role of HDL is complex,and under certain environmental stimulation,there may be both pro-inflammatory and anti-inflammatory effects.