LEI Xin-jun,MA Ai-qun,XI Yu-tao,et al.Expression of Kir2.1 channel during differentiation of human macrophages into foam cells[J].Journal of Southern Medical University,2005,(12):1461-1467.





Expression of Kir2.1 channel during differentiation of human macrophages into foam cells
雷新军1 马爱群1 席雨涛1 张葳2 姚艳3 杜媛1
1. 西安交通大学医学院第一附属医院心内科, 离子通道病研究室, 环境与疾病相关基因教育部重点实验室, 陕西, 西安, 710061;
2. 西安交通大学医学院第一附属医院儿科, 陕西, 西安, 710061;
3. 西安交通大学医学院第一附属医院呼吸科, 陕西, 西安, 710061
LEI Xin-jun1 MA Ai-qun1 XI Yu-tao1 ZHANG Wei2 YAO Yan3 DU Yuan1
1. Department of Cardiology, Ion Channelopathy Laboratory, Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, First Affiliated Hospital of School of Medicine, Xi’an Jiaotong University, Xi’an 710061, China;
2. Department of Pediatrics, First Affiliated Hospital of School of Medicine, Xi’an Jiaotong University, Xi’an 710061, China
ion channelcholesterol metabolismcell differentiationatherosclerosis
目的 Kir2.1通道是控制骨髓源性巨噬细胞增殖、活化和凋亡的重要离子通道之一,并且还参与细胞分化。本研究观察Kir2.1通道mRNA及蛋白在人单核细胞源性巨噬细胞向泡沫细胞分化过程中的表达。方法 采用密度梯度离心加贴壁黏附法,从男性健康志愿者的外周血中分离单核细胞,经5d培养后分化为巨噬细胞。在建立人巨噬细胞源性泡沫细胞模型的基础上,采用RT-PCR,蛋白质印迹及免疫细胞化学方法研究Kir2.1通道的表达。结果 将巨噬细胞同30mg/L氧化修饰低密度脂蛋白(OxLDL)于37℃孵育60h后,细胞体积增大,并有许多红色的脂质颗粒沉积于细胞质内,细胞内的总胆固醇(TC)、游离胆固醇(FC)及胆固醇酯(CE)的含量分别从(54.79±28.304)mg/g、(47.968±26.787)mg/g和(6.822±3.437)mg/g增加至(229.775±57.453)mg/g、(96.241±24.003)mg/g和(133.535±36.292)mg/g,CE/TC从(14.437±6.781)%提高到[(57.946±3.507)%(n=7,P<0.05)]。但是,Kir2.1通道mRNA的扩增量在巨噬细胞和分化的泡沫细胞之间无显著差异[(59.074±10.566)%vs(46.98±12.527)%,n=5,P>0.05];同样,Kir2.1通道蛋白在巨噬细胞和分化的泡沫细胞之间亦无显著差异[(60.527±18.621)%vs(50.243±11.583)%,n=6,P>0.05]。结论 人单核细胞源性巨噬细胞同30mg/LOxLDL孵育60h后可分化为泡沫细胞,但Kir2.1通道的表达无明显改变。
Objective Detected in non-transformed bone marrow-derived macrophages (BMDM) and identified as one of the key channels in modulating macrophage proliferation, activation and apoptosis, Kir2.1 channel is also characterized to play a crucial role in cell differentiation. The purpose of this study was to investigate the expression of Kir2.1 channel mRNA and protein during human monocyte-derived macrophage differentiation into foam cells. Methods Human peripheral blood monocytes were isolated from healthy male volunteers by density gradient centrifugation and then by adherence method. The macrophages identified as a homogeneous population of adherent cells were obtained after 5 days of culture. Expression of Kir2.1 channel during human macrophage differentiation into foam cells was investigated by RT-PCR, Western blotting and immunocytochemistry, respectively. Results After incubation of the macrophages with 30 mg/L OxLDL at 37℃ for 60 h, the cells were obviously enlarged in size and numerous red lipid granules observed under optical microscope. The cellular contents of the total cholesterol (TC), free cholesterol (FC) and cholesterol ester (CE) were markedly increased from 54.79±28.304 mg/g, 47.968±26.787 mg/g and 6.822±3.437 mg/g to 229.775±57.453 mg/g, 96.241±24.003 mg/g and 133.535±36.292 mg/g, respectively; the CE/TC ratio rose from (14.437±6.781)% to (57.946±3.507)% (n=7, P&lt;0.05), suggesting the phenotype of foam cells. However, there was no significant difference in the relative expression of Kir2.1 channel mRNA between the macrophages and foam cells [(59.074±10.566)% vs (46.98±12.527)%, n=5, P>0.05], nor was there significant difference in the relative expression of Kir2.1 channel protein between them [(60.527±18.621)% vs (50.243±11.583)%, n=6, P>0.05]. <b>Conclusion</b> Incubation of human monocyte-derived macrophages with 30 mg/L OxLDL for 60 h induces the differentiation of the cells into foam cells, but the expression of Kir2.1 channel does not change obviously.


[1] Jaber J,Murin J,Kinova S,et al.The role of infection and inflammation in the pathogensis of atherosclerosis[J].Vnitr Lek,2002,48(7):657-6.
[2] van Berkel TJ,Van Eck I,Herijgers N,et al.Scavenger receptor class A and B.Their role in atherogenesis and the matabolism of modified LDL and HDL[J].Ann N Y Acad Sci,2000,902:113-26.
[3] Antonov AS,Kolodgie FD,Munn DH,et al.Regulation of macrophage foam cell formation by alphaVbeta3 integrin:potential role in human atherosclerosis[J].Am J Pathol,2004,165(1):247-58.[4] Hakkinen T,Karkola K,Yla-Herttuala S.Macrophages,smooth muscle cells,endothelial cells,and T-cells expression CD40 and CD40L in fatty streaks and more advanced human atherosclerotic lesions.Colocalization with epitopes of oxidized low-density lipoprotein,scavenger receptor,and CD 16 (Fc gamma R Ⅲ) [J].Virchows Arch,2000,437(4):396-405.
[5] Gallin EK.Ion channels in leukocytes[J].Physiol Rev,1991,71(3):775-811.
[6] Eder C.Ion channels in microglia (brain macrophages) [J].Am J Physiol,1998,275(2 Pt 1):C327-C42.
[7] Soler C,Garcia-Manteiga J,Valdes R,et al.Macrophages require different nucleoside transport systems for proliferation and activation[J].FASEB J,2001,15(11):1979-88.
[8] Vicente R,Escalada A,Coma M,et al.Differential voltagedependent K+ channel responses during proliferation and activation in macrophages [J].J Biol Chem,2003,278(47):46307-20.
[9] Dallaporta B,Marchetti P,de Pablo MA,et al.Plasma membrane potential in thymocyte apoptosis [J].J Immunol,1999,162 (11):6534-42.
[10] Bernheim L,Bader CR.Human myoblast differentiation:Ca2+ channels are activated by K+ channels [J].News Physiol Sci,2002,17:22-6.
[11] Konig S,Hinard V,Arnaudeau S,et al.Membrane hyperpolarization triggers myogenin and myocyte enhancer factor-2 expression during human myoblast differentiation [J].J Biol Chem,2004,279 (27):28187-96.
[12] Ares MP,Kallin B,Eriksson P,et al.Oxidized LDL induces transcription factor activator protein-1 but inhibits activation of nuclear factor-kB in human vascular smooth muscle cells [J].Arterioscler Thromb Vasc Biol,1995,15(10):1584-90.
[13] Hirano K,Yamashita S,Nakagawa Y,et al.Expression of human scavenger receptor class B type I in cultured human monocytederived macrophages and atherosclerotic lesions [J].Circ Res,1999,85(1):108-16.
[14] Lei XJ,Ma AQ,Ren BW,et al.Fucoidin inhibits oxidized low density lipoprotein from inducing human peripheral blood monocyte expression of proinflammatory cytokines mRNA [J].ACAD J XJTU,2003,15(1):71-4.
[15] Hamilton JA,Whitty G,Jessup W.Oxidized LDL can promote human monocyte survival [J].Arterioscler Thromb Vasc Biol,2000,20(10):2329-31.
[16] Hamilton JA,Myers D,Jessup W,et al.Oxidized LDL can induce macrophage survival,DNA synthesis,and enhanced proliferative response to CSF-1 and GM-CSF [J].Arterioscler Thromb Vasc Biol,1999,19(1):98-105.
[17] Fach EM,Garulacan LA,Gao J,et al.In vitro biomarker discovery for atherosclerosis by proteomics [J].Mol Cell Proteomics,2004,3(12):1200-10.
[18] Romanenko VG,Fang Y,Byfield F,et al.Cholesterol sensitivity and lipid raft targeting of Kir2.1 channels[J].Biophys J,2004,87(6):3850-61.
[19] Fischer-Lougheed J,Liu JH,Espinos E,et al.Human myoblast fusion requires expression of functional inward rectifier Kir2.1channels[J].J Cell Biol,2001,153(4):677-86.
[20] Freedman BD,Price MA,Deutsch CJ.Evidence for voltage modulation of IL-2 production in mitogen-stimulated human peripheral blood lymphocytes[J].J Immunol,1992,149(12):3784-94.
[21] Cahalan MD,WulffH,Chandy KG.Molecular properties and physiological roles of ion channels in the immune system [J].J Clin Immunol,2001,21(4):235-52.
[22] Berridge MJ.Inositol trisphosphate and calcium signaling [J].Nature,1993,361(6410):315-25.


 WANG Jun-kui,CUI Chang-cong,YAO Qing-hai,et al.Na+/Ca2+ exchanger current and K+ current remodeling in midmyocardial cells of hypertrophic left ventricle[J].Journal of Southern Medical University,2004,(12):430.
[3]雷新军,张葳,林显丰,等.Diclofenac inhibits Kv1.3 and Kir2.1 expressions in human macrophages and affects the membrane potential and foam cell formation[J].南方医科大学学报,2012,(08):1067.
 LEI Xinjun,ZHANGWei,LIN Xianfeng,et al.双氯芬酸对人巨噬细胞钾通道Kv1.3、Kir2.1表达的抑制作用及其对膜电位和泡沫细胞形成的影响[J].Journal of Southern Medical University,2012,(12):1067.


基金项目:This work was supported by China Medical Board of New York, Inc.(#01-761) and Key International Science & Technology Cooperation Projects from Ministry of Science and Technology (2003DF000037),China.
更新日期/Last Update: