CAJ | 학술논문

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IFN 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。
목적：탐토 M1화 M2형거서세포경양화형저밀도지단백(ox LDL)자격후세포내지질함량급분비적세포인자백세포개소10(IL 10)、종류배사인자(TNF α)적농도차별급의의。방법제취 C57BL/6소서적골수단핵세포,용거서세포집락자격인자(M CSF)유도기향거서세포분화,병보이간우소γ(IFN γ)화백세포개소4(IL 4)자격거서세포향M1화M2형분화,행RT PCR검측겁화후 M1형거서세포적표지물일양화담합성매(iNOS)급 M2형거서세포적표지물정안산매(Arg1)적표체。M1급 M2형거서세포유도분화성공후재40 mg/L ox LDL중배육48 h후행유홍O염색관찰세포내지질함량；행ELISA검측상청액중IL 10화TNF α적농도。결과 RT PCR결과현시Arg1적표체재IFN γ자격조저우IL 4자격조,이iNOS적표체재IFN γ자격조고우IL 4자격조,표명 IFN γ자격후거서세포분화위 M1형이 IL 4자격후분화위 M2형；유홍 O염색시 M1화 M2형거서세포내적지질함량균고우대조조,M1형거서세포내지질함량고우 M2형거서세포。ELISA검측시 M1화M2형거서세포상청액중TNF α화IL 10농도균고우대조조,차이유통계학의의(P<0.05)；M1형거서세포상청액중 IL 10농도저우 M2형거서세포,이 TNF α적농도칙고우 M2형거서세포,차이유통계학의의(P<0.05)。결론 M1형거서세포가능통과분비 TNF α증강대담고순적섭입,가속동맥죽양경화적발생발전,이 M2형거서세포가능통과분비 IL 10감소대담고순적탄서,종이발휘항 AS작용。
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 macrophages 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 interferon 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.