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《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:

2012年第5期

页码:

559-564

栏目:

基础研究

出版日期:

2012-10-25

文章信息/Info

Title:

Developmental change of MaxiK channel during human macrophage-derived foam cell differentiation

作者:

雷新军¹; 张 葳²; 马爱群¹; 3; 袁祖贻¹; 3

(西安交通大学：1.医学院第一附属医院心内科，
2.医学院第一附属医院新生儿科，
3.环境与疾病相关基因教育部重点实验室，陕西 西安 710061)

Author(s):

LEI Xin-jun¹;  ZHANG Wei²;  Ma Ai-qun¹; 3;  Yuan Zu-yi¹; 3

(1.Department of Cardiology, 2.Department of Neonatal Neonatology, First Affiliated Hospital, Medical School, 3.Research Laboratory of Ion Channelopathy, Ministry of Education Key Laboratory of Environment and Gene-related Diseases, Xi’an Jiaotong University, Xi’an 710061, Shaanxi, China)

关键词:

膜片钳技术; 离子通道; 巨噬细胞; 细胞分化; 动脉粥样硬化

Keywords:

patch clamp technique;  ion channels;  macrophages;  cell differentiation;  atherosclerosis

分类号:

R363.2

DOI:

-

文献标识码:

A

摘要:

目的:研究人巨噬细胞发育及其向泡沫细胞分化过程中MaxiK通道的表达和电生理学特征。方法： 以健康人外周血单核细胞源性巨噬细胞为研究对象，采用Real time RT-PCR和Western blot技术观察MaxiK通道α亚单位mRNA及蛋白的表达；膜片钳技术分析MaxiK通道的电生理学特征。结果： 在巨噬细胞发育过程中，MaxiK通道α亚单位的表达被轻度上调。同培养5 d的细胞相比，7.5 d细胞的mRNA及蛋白表达增加分别约为1.06和1.44倍，但无统计学意义。然而，30 mg/L oxLDL显著提高MaxiK通道α亚单位的表达，在其分化成泡沫细胞后，mRNA和蛋白表达分别是培养5 d细胞的2.4和7.27倍，有显著的差异（P<0.05）。在所有培养5 d、7.5 d和oxLDL组中的巨噬细胞上均能记录到典型的MaxiK电流；MaxiK通道的选择性阻断剂- paxilline（10 μmol/L）抑制时间依从性电流、几乎全部的外向电导和噪声；但是，在培养5 d、7.5 d和oxLDL组中的巨噬细胞上MaxiK电流密度分别是(36±6) pA/pF、(35.9±3.5) pA/pF和(32.4±6.9) pA/pF，无明显差异。结论： 在人巨噬细胞发育过程中，MaxiK通道的表达被上调，分化成泡沫细胞后尤为显著，但其介导的电流没有改变。

Abstract:

AIM:To study the expression and electrophysiological characteristics of MaxiK channels in the process of human macrophage development and differentiation to foam cells. METHODS: Monocyte-derived macrophages were isolated from human peripheral blood. The mRNA level and MaxiK channel expression were detected using real time RT-PCR and Western blot technique and the whole cell MaxiK current from macrophages was measured using patch clamp technique. RESULTS: In the development of macrophage cells, the expression of MaxiK channel was slightly upregulated. Compared with those in the 5-day group, mRNA and protein expression in the 7.5-day group increased 1.06 and 1.44 times, respectively. However, 30 mg/L oxLDL significantly increased the mRNA expression of MaxiK. After differentiating into foam cells, MaxiK mRNA and protein expression were 2.4 and 7.27 times higher than those in the 5-day group (P<0.05). The MaxiK current was obtained from 5 day, 7.5 day and oxLDL groups. Paxilline (10 μmol/L), the selective blocker of MaxiK, inhibited the time-dependent current and abolished the outward conductance and noises. In 5-day, 7.5-day and oxLDL groups, the MaxiK current densities were (36±6) pA/pF, (35.9±3.5) pA/pF and (32.4±6.9) pA/pF respectively, Which did not show a significant difference. CONCLUSION: In the development of human macrophages, expression of MaxiK channel is upregulated. Upregulation is more obvious after its differentiation into foam cells. However, the MaxiK current remains unchanged during development and differentiation.

参考文献/References

[1]McLaren JE,Michael DR,Ashlin TG,et al.Cytokines, macrophage lipid metabolism and foam cells:implications for cardiovascular disease therapy[J].Prog Lipid Res,2011,50(4):331-47.
[2]喻 红.巨噬细胞-动脉粥样硬化的治疗靶点[J].中国动脉硬化杂志,2009,17(7):619-619.
[3]Blunck R,Scheel O,Müller M,et al.New insights into endotoxin-induced activation of macrophages: involvement of a K+ channel in transmembrane signaling[J].J Immunol,2001,166(2):1009-1015.
[4]Schilling T,Eder C.Lysophosphatidylcholine- and MCP-1-induced chemotaxis of monocytes requires potassium channel activity[J].Pflugers Arch,2009,459(1):71-77.
[5]Papavlassopoulos M,Stamme C,Thon L,et al.MaxiK blockade selectively inhibits the lipopolysaccharide-induced I kappa B-alpha/NF-kappa B signaling pathway inmacrophages[J].J Immunol,2006,177(6):4086-4093.
[6]雷新军,马爱群,席雨涛,等.阻断MaxiK通道对人单核细胞源性巨噬细胞向泡沫细胞分化的抑制作用[J].细胞与分子免疫学杂志,2006,22(3):310-314.
[7]Hansson GK,Hermansson A.The immune system in atherosclerosis[J].Nat Immunol,2011,12(3):204-212.
[8] Insull W Jr.The pathology of atherosclerosis: plaque development and plaque responses to medical treatment[J].Am J Med,2009,122(1 Suppl):S3-S14.
[9]Sprague AH,Khalil RA.Inflammatory cytokines in vascular dysfunction and vascular disease[J].Biochem Pharmacol,2009,78(6):539-552.
[10]Van Berkel TJ,Van Eck M,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,2000,902:113-126.
[11]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):247-258.
[12]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 CD16 (Fc gamma RⅢ)[J].Virchows Arch,2000,437(4):396-405.
[13]Papavlassopoulos M,Stamme C,Thon L,et al.MaxiK blockade selectively inhibits the lipopolysaccharide-induced I kappa B-alpha/NF-kappa B signaling pathway inmacrophages[J].J Immunol,2006,177(6):4086-4093.
[14]谭健苗，杨永宗，任 为，等.泡沫细胞形成与胞浆Ca2+水平变化[J].中国病理生理杂志,2002,18(2):131-135.
[15]Toro B,Cox N,Wilson RJ,et al.KCNMB1 regulates surface expression of a voltage and Ca2+-activated K+ channel via endocytic trafficking signals[J].Neuroscience,2006,142(3):661-669.

备注/Memo

备注/Memo:

收稿日期：2012-06-15.
基金项目：国家自然科学基金资助（81170276）；陕西省自然基金资助（2009JM4006）；西安交通大学医学创新基金资助（JH0203111）
通讯作者：袁祖贻，主任医师，主要从事动脉粥样硬化发病机制研究Email:zuyiyuan@mail.xjtu.edu.cn
作者简介：雷新军，主治医师，博士Email:xinjunlei@sohu.com
共同第一作者：张葳，主治医师，博士生Email:zhangweilei@sohu.com

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更新日期/Last Update: 2012-11-16