M1和M2型巨噬细胞经ox LDL刺激后细胞内脂质含量及分泌TNF α、IL 10的浓度差别与意义

M1和M2型巨噬细胞经ox LDL刺激后细胞内脂质含量及分泌TNF α、IL 10的浓度差别与意义
尚茹茹;张锦;袁锋;温婷;刘晓红
【期刊名称】《中西医结合心脑血管病杂志》
【年(卷),期】2014(000)012
【摘要】目的：探讨 M1和 M2型巨噬细胞经氧化型低密度脂蛋白(ox LDL)刺激后细胞内脂质含量及分泌的细胞因子白细胞介素10(IL 10)、肿瘤坏死因子(TNF α)的浓度差别及意义。

方法提取 C57BL/6小鼠的骨髓单核细胞,用巨噬细胞集落刺激因子(M CSF)诱导其向巨噬细胞分化,并辅以干扰素γ(IFN γ)和白细胞介素4(IL 4)刺激巨噬细胞向M1和M2型分化,行RT PCR检测极化后 M1型巨噬细胞的标志物一氧化氮合成酶(iNOS)及 M2型巨噬细胞的标志物精氨酸酶(Arg1)的表达。

M1及 M2型巨噬细胞诱导分化成功后在40 mg/L ox LDL中培育48 h后行油红O 染色观察细胞内脂质含量；行ELISA检测上清液中IL 10和TNF α的浓度。

结果RT PCR结果显示Arg1的表达在IFN γ刺激组低于IL 4刺激组,而iNOS的表达在IF N γ刺激组高于IL 4刺激组,表明IFN γ刺激后巨噬细胞分化为 M1型而 IL 4刺激后分化为 M2型；油红 O染色示 M1和 M2型巨噬细胞内的脂质含量均高于对照组,M1型巨噬细胞内脂质含量高于 M2型巨噬细胞。

ELISA检测示 M1和M2型巨噬细胞上清液中TNF α和IL 10浓度均高于对照组,差异有统计学意义
(P<0.05)；M1型巨噬细胞上清液中 IL 10浓度低于 M2型巨噬细胞,而TNF α的浓度则高于 M2型巨噬细胞,差异有统计学意义(P<0.05)。

结论 M1型巨噬细胞可能通过分泌TNF α增强对胆固醇的摄入,加速动脉粥样硬化的发生发展,而 M2型巨噬细胞可能通过分泌 IL 10减少对胆固醇的吞噬,从而发挥抗 AS作
用。

%Objective To investigate the differences and significance of intracel ular lipid contents in M1 and M2 macrophages and concentrations of TNF αand IL 10 secreted by M1 and M2 macrophage s after stimulating with oxidized low density lipopro-tein(ox LDL).Methods Monocytes were isolated from bone barrow cel s of C57BL/6 murine and were induced into macrophages by M CSF.Macrophages randomly al ocated into two groups and induced with interfero n gamma(IFN γ)and Interleukin 4(IL 4)to establish M1 and M2 macrophages,respectively.RT PCR was used to quantitative analysis Arg 1 expression which is M1 macro-phages marker and iNOS expression which is M2 macrophages marker.Then M1 and M2 macrophages were incubated with 40mg/L ox LDL for 48 h,accumulation of lipid was quantid by Oil Red O staining and the concentrations of cytokine IL 10 and TNF αin cel culture supernatants were measured by ELISA.Results Expression levels of Arg1 measured by RT PCR were lower in IFN
γstimulated macrophages(M1 macrophages)than IL 4 stimulated macrophages(M2 macrophages).Conversely,expression levels of iNOS was higher in IFN γstimulated macrophages. Lipid content in M1 and M2 macrophages was higher than those in control group,lipid content in M1 macrophages was higher than M2 macrophages.ELISA results indicate that supernatant concentrations of TNF αand IL 10 were higher than those in control group,the supernatant concentrations of IL 10 in M1 macrophages was lower than M2 macrophages(P<0.05).In contrast,the supernatant concentration of TNF αin M1 macrophages was higher than M2 mac-rophages(P<0.05).Conclusion M1 macrophages possibly increase the
ability of cholesterol intake by secretion of TNF αand play the role in proatherogenic functions,while M2 macrophages may reduce cholesterol phagocytosis by secretion of IL 10 and play the role of antiatherogenic functions.
【总页数】3页(P1537-1539)
【作者】尚茹茹;张锦;袁锋;温婷;刘晓红
【作者单位】山西医科大学第一临床医学院太原 030001;山西省人民医院太原030001;山西医科大学太原 030001;山西医科大学太原 030001;山西省人民医院太原 030001
【正文语种】中文
【中图分类】R541;R256.2
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