核苷酸结合寡聚结构域受体1作为先天免疫识别受体在血管平滑肌细胞激活中的作用★
丁彦春，曲 鹏
课题背景：核苷酸结合的寡聚结构域受体是新近发现的细胞内模式识别受体，目前的研究主要集中于它们在人类一些炎症性疾病如炎症性肠病的发生、发展中的作用。核苷酸结合的寡聚结构域受体与Toll样受体具有相似的结构和功能，核苷酸结合的寡聚结构域受体是否像Toll样受体一样也参与动脉粥样硬化形成和发展，它们在动脉粥样硬化形成和发展中的具体作用机制，以及二者介导的信号传导通路之间的相互作用关系将是未来该领域的研究热点。
同行评价：文章通过观察细菌细胞壁粘肽对血管平滑肌中细胞内模式识别受体核苷酸结合的寡聚结构域受体1激活和表达的影响，结论认为核苷酸结合的寡聚结构域受体1介导的先天免疫信号通路可能通过激活血管平滑肌细胞参与动脉粥样硬化的发生和发展。对于临床上预防动脉硬化形成和经皮冠状动脉介入治疗术后再狭窄过程有重要意义。
相关链接：免疫反应参与动脉粥样硬化形成的全过程，先天免疫反应的激活在动脉粥样硬化形成中具有重要作用。对先天免疫反应及模式识别受体在动脉粥样硬化形成中的作用进行研究不仅有助于理解动脉粥样硬化形成过程，而且还能通过直接干扰动脉粥样硬化形成中的炎症反应过程来提供抗动脉粥样硬化治疗的新方案、新策略。
摘要
目的:核苷酸结合的寡聚结构域受体1 是新近发现的先天免疫细胞内模式识别受体。实验通过观察细菌细胞壁粘肽对血管平滑肌中细胞内核苷酸结合的寡聚结构域受体1激活和表达的影响，探讨核苷酸结合的寡聚结构域受体1介导的先天免疫信号通路在血管平滑肌激活中的作用。
方法：实验于2006-06/2007-03在大连医科大学附属二院中心实验室完成。①实验材料：人冠状动脉血管平滑肌细胞购自Cambrex公司。②实验过程：体外培养人冠状动脉血管平滑肌细胞，用核苷酸结合的寡聚结构域受体1激动剂细菌细胞壁粘肽（10 mg/L）刺激0，3，6和24 h。③实验评估：用实时定量反转录-聚合酶链反应检测平滑肌细胞中核苷酸结合的寡聚结构域受体1 mRNA的表达，用酶联免疫吸附法检测细胞培养物上清液中白细胞介素8和肿瘤坏死因子α的浓度。
结果：①血管平滑肌细胞在静息状态下表达低水平的核苷酸结合的寡聚结构域受体1。细菌细胞壁粘肽刺激能够引起血管平滑肌细胞中核苷酸结合的寡聚结构域受体1 mRNA表达上调(从0.164±0.005增加到0.231±0.027，P < 0.05)。②酶联免疫吸附法显示细胞培养物上清液中白细胞介素8和肿瘤坏死因子α的浓度也显著增加[白细胞介素8从（0.12±0.01）μg/L 增加到（2.31±0.11）μg/L，肿瘤坏死因子α从（4.22±0.50）ng/L 增加到（143.11±12.58）ng/L，P < 0.01]。
结论：推测血管平滑肌细胞中的核苷酸结合的寡聚结构域受体1介导的先天免疫信号通路被激活，参与了动脉粥样硬化的发生和发展。
关键词：血管平滑肌细胞；核苷酸结合的寡聚结构域受体1；粘肽；动脉粥样硬化；血管组织构建����
丁彦春，曲鹏. 核苷酸结合寡聚结构域受体1作为先天免疫识别受体在血管平滑肌细胞激活中的作用[J].中国组织工程研究与临床康复，2008，12(11):2021-2024 [www.zglckf.com/zglckf/ejournal/upfiles/08-11/11k-2021(ps).pdf]
大连医科大学附属第二医院心内科，辽宁省大连市 116027
丁彦春★，男，1971年生，河南省扶沟县人，汉族，1999年大连医科大学毕业，硕士，副教授，主要从事动脉硬化形成机制及介入心脏病学。
yanchunding@
yahoo.com.cn
通讯作者：曲 鹏，博士，教授，大连医科大学附属第二医院心内科，辽宁省大连市
116027 qupeng777@
yahoo.com.cn
中图分类号:R329.412
文献标识码:A
文章编号:1673-8225
(2008)11-02021-04
收稿日期：2007-12-21
修回日期：2008-02-07 (07-50-12-7097/W·Y)
Role of nucleotide binding oligomerization domain receptor 1 as a new innate immune recognition receptor in activation of vascular smooth muscle cells
Abstract
AIM：Nucleotide binding oligomerization domain receptor 1 (NOD1) is a recently identified intracellular pathogen pattern recognition receptor of innate immunity. This study was designed to explore the role of NOD1-mediated innate immune signal pathway in the activation of vascular smooth muscle cells (VSMCs), and investigate the effect of peptidoglycan (PGN) on the activation and expression of NOD1 in human VSMCs.
METHODS: The experiment was performed in the central laboratory of the Second Affiliated Hospital of Dalian Medical University from June 2006 to March 2007.①Subject: Human coronary artery VSMCs were purchased from Cambrex company.②Methods: Human coronary artery VSMCs were cultured in vitro, and stimulated with NOD1 agonist PGN (10 mg/L) for 0, 3, 6 and 24 hours.③Evaluation: The mRNA expression of NOD1 in VSMCs was measured by real time quantitative reverse transcription-polymerase chain reaction. The concentration in the culture supernatants of interleukin-8 (IL-8) and tumor necrosis factor-α (TNF-α) was determined by enzyme linked immunosorbent assay.
RESULTS: ①Human VSMCs constitutively expressed a low level of NOD1 at resting condition. Upon PGN stimulation, the expression of NOD1 mRNA was up-regulated in VSMCs, from 0.164±0.005 to 0.231±0.027 (P < 0.05).②Enzyme linked immunosorbent assay showed the concentration in the culture supernatants of IL-8 and TNF-α increased significantly, IL-8 from (0.12±0.01) μg/L to (2.31±0.11) μg/L and TNF-α from (4.22±0.50) ng/L to (143.11±12.58) ng/L, respectively (P < 0.01).
CONCLUSION: The NOD1-mediated innate immune pathway may be involved in the atherogenesis though the activation of VSMCs.
Ding YC, Qu P. Role of nucleotide binding oligomerization domain receptor 1 as a new innate immune recognition receptor in activation of vascular smooth muscle cells.Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu 2008;12(11):2021-2024(China) [www.zglckf.com/zglckf/ejournal/upfiles/08-11/11k-2021(ps).pdf]
Department of Cardiology, Second Affiliated Hospital of Dalian Medical University, Dalian 116027, Liaoning Province, China
Ding Yan-chun★, Master, Associate professor, Department of Cardiology, Second Affiliated Hospital of Dalian Medical University, Dalian 116027, Liaoning Province, China
yanchunding@
yahoo.com.cn
Correspondence to: Qu Peng, Doctor, Professor, Department of Cardiology, Second Affiliated Hospital of Dalian Medical University, Dalian 116027, Liaoning Province, China
qupeng777@
yahoo.com.cn
Received: 2007-12-21
Accepted: 2008-02-07
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