ADMA对单核细胞来源的树突状细胞成熟和免疫的影响(1)
中图分类号：R446 文献标 识码：A 文章编号：1009_816X(2010)06_0428_04
DOI:10.3969/j.issn.1009_816X.2010.06.08 Effect of Asymmetrical Dimethylarginine(ADMA)on Maturation and Function of Den dritic Cells Drived from Monocyte.XU Hua_bao, ZHU Wei_guo, LI Shan, et al. Zhuji People's Hospital, Zhejiang 31180 0,China
[Abstract] ObjectiveThe purpose of this study is to explore the mechanism of ge neration and progress of AS by investigating the effect of ADMA on the expressio n of dendritic cells(DCs) and the interaction between DCs and T lymphocytes.MethodsImmunophenotype, such as HLA_DR, CD1a, CD86 and CD83, of ADMA_treated DCswas monitored by flow cytometry. Allogeneic T cell activation by ADMA_treated DC s was tested by mixed lymphocyte reaction (MLR). The apoptosis of ADMA_treated D Cs was measured by flow cytometry using a Annexin V_FITC. The expression of IL_ 10, IL_12 and TNF_α on ADMA_treated DCs was detected by immunofluorescence sta in.Results The majority of immature DCs cultured in control cultures with GM_C SF and IL_4 expressed high level of HLA_DR, moderate levels of CD1a and CD86, a n d low level of CD83, while the mature DCs exposed to LPS expressed high levels o f HLA_DR, CD86 and CD83, moderate level of CD1a. The presence of ADMA did not c hange the phenotype of immature DCs and high concentration ADMA inhibit the matu ration of DCs. Immature DCs in high concentration ADMA were less effective in st imulating T_cell proliferation. And immature DCs in high concentration ADMA weremore effective to apoptosis. The expression of IL_10, IL_12 and TNF_α on DCs w as significantly reduced by ADMA in a concentration_dependent manner as assessedby immunofluorescence stain.ConclusionsHigh concentration ADMA is able to inhibit the maturatiom of DCs and promote theapoptosis of DCs. ADMA can reduce the expression of IL_10, IL_12 and TNF_α onDCs. These data indicate a novel pathophysiological mechanism for ADMA promotin g atherogenesis.
, 百拇医药
[Key words] Asymmetrical dimethylarginine(ADMA); Dendritic c ells(DCs); Atherosclerosis(AS); In vitro test
近年来，越来越多的证据表明动脉粥样硬化(atherosclerosis，AS)是一种慢性炎症和自 身免疫性疾病[1]。树突状细胞(dendritic cells，DCs)是目前体内功能最强大 的抗原递呈细胞(antigen presenting cells，APCs)[2]，它们在外周组织中监 控入侵的病原体，捕获和处理抗原，并递呈给初始型T淋巴细胞，使之激活、增殖和分化， 从而介导机体产生相应的免疫应答[3]。最近，发现DCs存在于正常动脉壁中，而在 粥样斑块中数量明显增多，说明DC可能在致AS的炎症反应中起重要作用[4]。内源 性非对称性二甲基精氨酸(ADMA)对一氧化氮合酶(NOS)具有强效抑制作用，健康人血浆AD MA水平大约是1umol/L，但这种血浆浓度水平不足以抑制NO合成，试验研究报道ADMA浓度在 病理生理高浓度范围(3～15umol/L)可明显抑制NO生物合成[4]。研究证实ADMA 参与AS的病理过程[4]，本文主要是研究不同浓度ADMA对DC的影响，从而为ADMA参 与AS的病理过程提供更多的理论基础。
, 百拇医药
1 材料和方法
1.1 主要试剂和材料：人重组集落刺激因子(recombinant human granulocyte macropha ge_colony stimulating factor，rhGM_CSF)和人重组白介素_4(recombinant human int erleukin_4，rhIL_4)均购自美国R＆D公司，RPMI 1640培养基购自美国Gibco公司，HLA_D R_FITC、CD86_FITC、CD83_phycoerythrin(PE)、CD80_PE购自美国Caltag公司，人淋巴细 胞分离液购自中国天津灏洋生物制品科技有限公司，ADMA标准品购自美国Sigma公司。采用 美国Biosource生产的Human IL_12、Human IL_10、Human TNF_α试剂盒。
1.2 树突状细胞的分离培养：用贴壁法分离培养树突状细胞，第五天随机分成五组，分别 为空白对照组，LPS组(1umol/L)，1、8、16umol/L浓度ADMA组，每组细胞浓度为4×10 7/ml，每组约1ml，24小时后收集悬浮细胞。, http://www.100md.com(徐华宝 朱伟国 李闪 朱建华)
DOI:10.3969/j.issn.1009_816X.2010.06.08 Effect of Asymmetrical Dimethylarginine(ADMA)on Maturation and Function of Den dritic Cells Drived from Monocyte.XU Hua_bao, ZHU Wei_guo, LI Shan, et al. Zhuji People's Hospital, Zhejiang 31180 0,China
[Abstract] ObjectiveThe purpose of this study is to explore the mechanism of ge neration and progress of AS by investigating the effect of ADMA on the expressio n of dendritic cells(DCs) and the interaction between DCs and T lymphocytes.MethodsImmunophenotype, such as HLA_DR, CD1a, CD86 and CD83, of ADMA_treated DCswas monitored by flow cytometry. Allogeneic T cell activation by ADMA_treated DC s was tested by mixed lymphocyte reaction (MLR). The apoptosis of ADMA_treated D Cs was measured by flow cytometry using a Annexin V_FITC. The expression of IL_ 10, IL_12 and TNF_α on ADMA_treated DCs was detected by immunofluorescence sta in.Results The majority of immature DCs cultured in control cultures with GM_C SF and IL_4 expressed high level of HLA_DR, moderate levels of CD1a and CD86, a n d low level of CD83, while the mature DCs exposed to LPS expressed high levels o f HLA_DR, CD86 and CD83, moderate level of CD1a. The presence of ADMA did not c hange the phenotype of immature DCs and high concentration ADMA inhibit the matu ration of DCs. Immature DCs in high concentration ADMA were less effective in st imulating T_cell proliferation. And immature DCs in high concentration ADMA weremore effective to apoptosis. The expression of IL_10, IL_12 and TNF_α on DCs w as significantly reduced by ADMA in a concentration_dependent manner as assessedby immunofluorescence stain.ConclusionsHigh concentration ADMA is able to inhibit the maturatiom of DCs and promote theapoptosis of DCs. ADMA can reduce the expression of IL_10, IL_12 and TNF_α onDCs. These data indicate a novel pathophysiological mechanism for ADMA promotin g atherogenesis.
, 百拇医药
[Key words] Asymmetrical dimethylarginine(ADMA); Dendritic c ells(DCs); Atherosclerosis(AS); In vitro test
近年来，越来越多的证据表明动脉粥样硬化(atherosclerosis，AS)是一种慢性炎症和自 身免疫性疾病[1]。树突状细胞(dendritic cells，DCs)是目前体内功能最强大 的抗原递呈细胞(antigen presenting cells，APCs)[2]，它们在外周组织中监 控入侵的病原体，捕获和处理抗原，并递呈给初始型T淋巴细胞，使之激活、增殖和分化， 从而介导机体产生相应的免疫应答[3]。最近，发现DCs存在于正常动脉壁中，而在 粥样斑块中数量明显增多，说明DC可能在致AS的炎症反应中起重要作用[4]。内源 性非对称性二甲基精氨酸(ADMA)对一氧化氮合酶(NOS)具有强效抑制作用，健康人血浆AD MA水平大约是1umol/L，但这种血浆浓度水平不足以抑制NO合成，试验研究报道ADMA浓度在 病理生理高浓度范围(3～15umol/L)可明显抑制NO生物合成[4]。研究证实ADMA 参与AS的病理过程[4]，本文主要是研究不同浓度ADMA对DC的影响，从而为ADMA参 与AS的病理过程提供更多的理论基础。
, 百拇医药
1 材料和方法
1.1 主要试剂和材料：人重组集落刺激因子(recombinant human granulocyte macropha ge_colony stimulating factor，rhGM_CSF)和人重组白介素_4(recombinant human int erleukin_4，rhIL_4)均购自美国R＆D公司，RPMI 1640培养基购自美国Gibco公司，HLA_D R_FITC、CD86_FITC、CD83_phycoerythrin(PE)、CD80_PE购自美国Caltag公司，人淋巴细 胞分离液购自中国天津灏洋生物制品科技有限公司，ADMA标准品购自美国Sigma公司。采用 美国Biosource生产的Human IL_12、Human IL_10、Human TNF_α试剂盒。
1.2 树突状细胞的分离培养：用贴壁法分离培养树突状细胞，第五天随机分成五组，分别 为空白对照组，LPS组(1umol/L)，1、8、16umol/L浓度ADMA组，每组细胞浓度为4×10 7/ml，每组约1ml，24小时后收集悬浮细胞。, http://www.100md.com(徐华宝 朱伟国 李闪 朱建华)