脂多糖对肠黏膜微血管内皮细胞表达TLR2和TLR4的影响

doi:10.11924/j.issn.1000-6850.casb14120159

中国农学通报 ›› 2015, Vol. 31 ›› Issue (11): 72-77.doi: 10.11924/j.issn.1000-6850.casb14120159

所属专题：生物技术

脂多糖对肠黏膜微血管内皮细胞表达TLR2和TLR4的影响

王慧明,赵华,孙英健,胡格,穆祥,段慧琴

北京农学院,北京农学院,北京农学院,北京农学院,北京农学院,北京农学院

收稿日期:2014-12-23 修回日期:2015-03-12 接受日期:2015-03-13 出版日期:2015-05-06 发布日期:2015-05-06
通讯作者: 段慧琴
基金资助:
北京市优秀人才培养资助个人项目“TLR4介导的LPS信号转导”(PYZZ090416001326)；北京市属高等学校人才强教计划资助项目PHR(IHLB)“ET-1/NO平衡研究与信号转导”(201008423)；国家自然基金资助项目“小檗碱抗炎新机制——调节MD-2表达与功能”(2033202037)；北京农学院科技创新团队科研能力提升工程项目“几种中兽药抗病机理的研究”(KCT2014012)。

Effect of lipopolysaccharide on the expression of TLR2 and TLR4 in intestinal mucosa microvascular endothelial cells

Received:2014-12-23 Revised:2015-03-12 Accepted:2015-03-13 Online:2015-05-06 Published:2015-05-06

摘要/Abstract

摘要： 为研究TLR2、TLR4在脂多糖(lipopolysaccharide,LPS)致肠黏膜微血管内皮细胞(RIMECs)损伤中的作用，以体外培养的肠黏膜微血管内皮细胞为模型，用浓度为1、5、10μg/mL的LPS刺激RIMECs3、6、9、12、24h后，采用半定量RT-PCR法检测细胞表面TLR2和TLR4的表达情况；再以浓度为1、5、10μg/mL的LPS刺激RIMECs9h后，半定量RT-PCR法检测细胞TLR2和TLR4的表达情况。结果发现：与对照组比较，不同浓度LPS刺激均能引起RIMECs表面TLR2和TLR4mRNA表达增加。不同浓度LPS刺激不同时间后，RIMECs表面TLR2和TLR4mRNA表达基本均在9h达到峰值，且无时间依赖性。用LPS刺激RIMECs9h，5μg/mL的LPS对细胞表达TLR2mRNA作用最强且无剂量依赖性，10μg/mLLPS对细胞表达TLR4mRNA作用最强且呈剂量依赖性。TLR2和TLR4在LPS损伤RIMECs的过程中起到重要作用。

关键词: 野生百合, 野生百合, 花蕾, 再生植株

Abstract: The aim of the study was to investigate the effect of TLR2, TLR4 in injury of rat intestinal mucosa microvascular endothelial cells (RIMECs) induced by lipopolysaccharide (LPS). Rat intestinal mucosa microvascular endothelial cell in vitro was taken as a model, and different concentrations of LPS (1, 5, 10 μg/mL) were used to stimulate RIMECs for different time (3, 6, 9, 12, 24 h). Expressions of TLR2 and TLR4 on cell surface were detected by semi quantitative RT-PCR method. Then LPS at different concentrations (1, 5, 10 g/mL) were used to stimulate RIMECs for 9 h. Then expressions of TLR2 and TLR4 cells were detected by semi quantitative RT-PCR method. The results showed that expression of TLR2 and TLR4 increased when RIMECs were stimulated by LPS compared with the control group. Moreover, after stimulation with different concentrations of LPS for different time, expression of TLR2 mRNA and TLR4 mRNA basically reached the peak at 9 h, and the expression was time-independent. Expression of TLR2 mRNA was the highest when the concentration of LPS was 5 μg/mL after 9 h, and the expression was dose-independent. LPS at a concentration of 10 μg/mL stimulating RIMECs had the strongest effect in the expression of TLR4 mRNA, and was dose-dependent. The results demonstrated that TLR2 and TLR4 played an important role in the process of LPS damaging RIMECs.

王慧明,赵华,孙英健,胡格,穆祥,段慧琴. 脂多糖对肠黏膜微血管内皮细胞表达TLR2和TLR4的影响[J]. 中国农学通报, 2015, 31(11): 72-77.

参考文献

[1] Yu Li,Hongtao Liu,Qing-Song Xu,et al. Chitosan oligosaccharides block LPS-induced O-GlcNAcylation of NF-κB and endothelial inflammatory response[J]. Carbohydrate Polymers,2014,99：568-578.
[2] AN HuaZhang,QIAN Cheng,CAO XueTao,et al. Regulation of Toll-like receptor signaling in innate immunity[J]. Life Sciences, 2010,53(1):34-43.
[3] Nicholas J. Gay, Monique Gangloff, et al. Structure and function of Toll receptors and their ligands[J], Annual Review of biochemistry,2007,76:141-165.
[4] Jin Young Kang, Jie-Oh Lee, et al. Structural Biology of the Toll-like receptor family[J], Annual Review of biochemistry,2011,80:917-941.
[5] O’Neill LA, Bryant CE, Doyle SL. Therapeutic targeting of Toll-like receptors for infectious and inflammatory diseases and cancer[J]. Pharmacol. Rev, 2009. 61:177–97.
[6] 索占伟、穆祥等,大鼠肠粘膜微血管内皮细胞的体外培养[J],解剖学报,2005,3(2)：103-109.
[7] Yamamoto M, Sato S, Hemmi H, et al. Essential role for TIRAP inactivation of the signaling cascade shared by TLR2 and TLR4[J]. Nature, 2002,420(6913):324-329.
[8] Xiaoxia Li, Jinzhong Qin. Modulation of Toll-interleukin 1 receptor mediated signaling[J]. Mol Med, 2005,83(4):258-266.
[9] 李雅琳、郑海伦、王启之等,Toll样受体4、核因子-кB通路在溃疡性结肠炎发病机制中的应用[J],蚌埠医学院学报,2013,38(1): 16-19.
[10] Sook-Kyoung Heo, Hyun-Jeong Yun, et al. LPS induces inflammatory reponses in human aortic vascular smooth muscle cells via Toll-like receptor 4 expression and nitric oxide production[J], Immunology Letters, 2008,120:57-64.
[11] 徐淑凤、王平等,脂多糖诱导肺血管内皮细胞炎性损伤机制的探讨[J],中华结核和呼吸杂志,2011,11(34)：816-819.
[12] 李杰、董虹等,大鼠不部位内皮细胞的体外培养[J],北京农学院学报,2012,3(27)：29-31.
[13] 姚勇明、盛志勇等,我国创伤脓毒症基础研究进展[J],中华创伤杂志,2003,1(19)：9-12.
[14] 王国兴等,Toll样受体及其信号转导[J],细胞与分子免疫学杂志,2002,18(6):666-669.
[15] Tetsuya Matsuguchi,Tipayaratn Musikacharoen,et al. Gene Expressions of Toll-Like Receptor 2, but not Toll-Like receptor 4, is induced by LPS and inflammatory cytokines in mouse macrophages[J], The Journal of Immunology, 2000: 165: 5767–5772.
[16] 谭琰、邹开芳等,Expression and Implication of Toll-like receptor TLR2,TLR4 and TLR9 in Colonic Mucosa of Patients with Ulcerative Colitis[J]. Journal of Huazhong University of Science and Technology (Medical Sciences), 2014,34(5):785-790.
[17] YANG Qingwu,ZHU Peifang,WANG Zhengguo et al. Lipopolysaccharide upregulates the expression of Toll like receptor 4 in human vascular endothelial cells[J]. Chinese Medical Journal, 2002,115(2):286-289.
[18] 刘聪等,LPS对母兔子宫中不同部位TLR2、TLR4及其免疫相关细胞因子表达的影响[D],四川农业大学,2013,6：1-2.
[19] George H, Michael M, Robert E, et al. Counteracting interactions between lipopolysaccharide molecules with differential activation of toll-like receptor[J].Infect Immun, 2002,70(12):6658-6664.
[20] Osamu Takeuchi, Katsuaki Hoshino, et al. Differential Roles of TLR2 and TLR4 in Recognition of Gram-Negative and Gram-Positive Bacterial Cell Wall Components[J]. Immunity,1999,10(11):443-461.

脂多糖对肠黏膜微血管内皮细胞表达TLR2和TLR4的影响

Effect of lipopolysaccharide on the expression of TLR2 and TLR4 in intestinal mucosa microvascular endothelial cells

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 13

编辑推荐

Metrics

本文评价

关于本刊

作者中心

审者中心

在线期刊

联系我们

[1]	赵敬一,罗丹,张喜春. ProDH基因反义表达载体的遗传转化[J]. 中国农学通报, 2015, 31(26): 78-83.
[2]	徐茜余鸿燕徐燕黎华廖采琴. 重庆野生百合花蕾诱导再生植株技术研究[J]. 中国农学通报, 2014, 30(7): 121-125.
[3]	田英常红宇王昊王锦秀. 2个宁夏枸杞新品种花蕾物质代谢水平研究[J]. 中国农学通报, 2014, 30(22): 165-168.
[4]	曾祥琼杨治国朱照静黄先元黄晓丽陈俊意. 修剪对青蒿花蕾青蒿素含量的影响[J]. 中国农学通报, 2013, 29(7): 155-158.
[5]	曾祥琼江玲何荣会王易振景戌甘晓玲李世敏王抒邓步华唐全陈俊意. 遮阴对青蒿花蕾青蒿素含量的影响[J]. 中国农学通报, 2011, 27(2): 230-233.
[6]	梁美霞. 番茄子叶和下胚轴离体再生体系建立[J]. 中国农学通报, 2010, 26(6): 47-50.
[7]	薛义霞李亚灵温祥珍. 空气湿度对高温下番茄生殖生长的影响[J]. 中国农学通报, 2010, 26(15): 291-295.
[8]	穆春华,刘霞,于元杰,王金辉. 章丘大葱子叶再生体系建立及再生株倍性鉴定[J]. 中国农学通报, 2008, 24(8): 355-359.
[9]	潘竟丽,,曾幼玲,张富春. 新疆大叶紫花苜蓿(Medicago sativa.L)子叶、下胚轴、根的植株再生[J]. 中国农学通报, 2007, 23(6): 52-52.
[10]	陈娟,,潘开文,辜彬,王进闯,吴越华,万涛. 白术叶片再生植株和丛生芽快繁的研究[J]. 中国农学通报, 2006, 22(11): 65-65.
[11]	向地英,张延龙,郝瑞杰. 秦巴山区及毗邻地区野生百合性状描述[J]. 中国农学通报, 2006, 22(10): 97-97.
[12]	张素勤,邹志荣,耿广东,舒志强,王海波. 植物激素直接诱导非洲菊花蕾不定芽的研究[J]. 中国农学通报, 2005, 21(6): 292-292.
[13]	陆瑞菊,王亦菲,孙月芳,周润梅,黄剑华. 玉米小孢子培养再生植株的移栽及倍性评估[J]. 中国农学通报, 2005, 21(3): 84-84.