|本期目录/Table of Contents|

[1]韩文正,武虎,李晓艳,等.LPS与巨噬细胞中htra2的表达及相关性研究[J].浙江理工大学学报,2013,30(03):383-388.
 HAN Wenzheng,WU Hu,LI Xiaoyan,et al.Study of Expression and Relevance of htra 2 in Macrophage and LPS[J].Journal of Zhejiang Sci-Tech University,2013,30(03):383-388.
点击复制

LPS与巨噬细胞中htra2的表达及相关性研究()
分享到:

浙江理工大学学报[ISSN:1673-3851/CN:33-1338/TS]

卷:
第30卷
期数:
2013年03期
页码:
383-388
栏目:
(自科)生物与生命科学
出版日期:
2013-05-10

文章信息/Info

Title:
Study of Expression and Relevance of htra 2 in Macrophage and LPS
文章编号:
1673-3851 (2013) 03-0383-06
作者:
韩文正 武虎 李晓艳 李红艳 姚韵靓 郭科妮 周秀梅
1. 浙江理工大学生命科学学院新元医学与生物技术研究所, 杭州 310018; 2. 浙江大学医学院免疫研究所, 杭州 310058
Author(s):
HAN Wenzheng WU Hu LI Xiaoyan LI Hongyan YAO Yunliang GUO Keni ZHOU Xiumei
1. Xinyuan Institute of Medicine and Biotechnology, School of Life Sciences,Zhejiang Sci-Tech University, Hangzhou 310018, China; 2. Institute of Immunology, School of Medicine, Zhejiang University, Hangzhou 310058, China
关键词:
巨噬细胞 LPS VSV 免疫荧光 IL6
分类号:
R392
文献标志码:
A
摘要:
通过LPS(lipopolysaccharide)分别刺激树突状细胞和巨噬细胞,发现巨噬细胞中HtrA丝氨酸蛋白酶2(HtrA serine peptidase 2, htra2)基因的表达随着刺激时间的延长呈现下调的变化趋势,提示巨噬细胞中的htra2受到LPS的调控。为了进一步证实LPS刺激的特异性,实验建立VSV(vesicular stomatitis virus)病毒感染巨噬细胞模型,采用荧光定量PCR,免疫荧光方法分别从 htra2的蛋白水平和定位上研究 htra2 的变化与LPS刺激巨噬细胞产生效应的相关性;构建了 htra2 基因过表达的质粒pKH3 htra2 HA,采用ELISA,Western Blot及荧光定量PCR等方法,检测过表达 htra2 对巨噬细胞产生和分泌炎症因子IL 6(Interleukin 6)及IL 1b(Interleukin 1b)的影响。实验结果表明: htra2 仅在LPS刺激巨噬细胞的体系中受到调控,且过表达的 htra2 会上调巨噬细胞IL 1b和IL 6的表达水平。

参考文献/References:

[1]Janeway Jr C A, Medzhitov R. Innate immune recognition[J]. Annual Review of Immunology, 2002, 20(1): 197-216.
[2] Papadimitraki E D, Bertsias G K, Boumpas D T. Toll like receptors and autoimmunity: a critical appraisal[J]. Journal of Autoimmunity, 2007, 29(4): 310-318.
[3] Aderem A, Ulevitch R J. Toll like receptors in the induction of the innate immune response[J]. Nature, 2000, 406 : 782-787.
[4] Akira S, Tokeda K, Kaisho T, et al. Toll like receptors: critical proteins linking innate and acquired immunity[J]. Nature Immunology, 2001, 2(8): 675-80.
[5] Georgel P, Jiang Z, Kunz S, et al. Vesicular stomatitis virus glycoprotein G activates a specific antiviral Tolllike receptor 4 dependent pathway[J]. Virology, 2007, 362(2): 304-313.
[6] Suzuki Y, Imai Y, Nakayama H, et al. A serine protease, HtrA2, is released from the mitochondria and interacts with XIAP, inducing cell death[J]. Molecular Cell, 2001, 8(3): 613-621.
[7] Li Wenyu, Srinivasula S M, Chai Jijie, et al. Structural insights into the proapoptotic function of mitochondrial serine protease HtrA2/Omi[J]. Nature Structural Biology, 2002, 9(6): 436-441.
[8] Martins L M, Iaccarino I, Tenev T, et al. The serine protease Omi/HtrA2 regulates apoptosis by binding XIAP through a reaperlike motif[J]. The Journal of Biological Chemistry, 2002, 277(1): 439-444.
[9] Li B, Hu Q, Wang H, et al. Omi/HtrA2 is a positive regulator of autophagy that facilitates the degradation of mutant proteins involved in neurodegenerative diseases[J]. Cell Death and Differentiation, 2010, 17: 1773-1784
[10] Lin H T, Tsai H Y, Liu C P, et al. Comparability of bovine virus titers obtained by TCID50/mL and FAID50/mL[J]. Journal of Virological Methods, 2010, 165(1): 121-124.
[11] Ornberg R L, Harper T F, Liu H, et al. Western blot analysis with quantum dot fluorescence technology: a sensitive and quantitative method for multiplexed proteomics[J]. Nature Methods, 2005, 2(1): 79-81.
[12] Hegde R, Srinivasula S M, Zhang Z J, et al. Identification of Omi/HtrA2 as a mitochondrial apoptotic serine protease that disrupts inhibitor of apoptosis proteincaspase interaction[J].The Journal of Biological Chemistry, 2002, 277(1): 432-438.
[13] Wright S D, Ramos R A, Tobias P S, et al. CD14, a receptor for complexes of lipopolysaccharide(LPS) and LPS binding protein[J]. Science, 1990, 249(4975): 1431-1433.
[14] Wang D, Lou J, Ou Y, et al. Rasrelated protein Rab10 facilitates TLR4 signaling by promoting replenishment of TLR4 onto the plasma membrane[J]. Proc Natl Acad Sci USA, 2010, 107(31): 13806-13811.
[15] Ndlovu N, Van Lint C, Van Wesemael K, et al. Hyperactivated NFκB and AP1 transcription factors、promote highly accessible chromatin and constitutive transcription across the interleukin6 gene promoter in metastatic breast cancer cells[J]. Molecular and Cellular Biology, 2009, 29(20): 5488-5504.
[16] Narayan O, Sheffer D, Clements J E, et al. Restricted replication of lentiviruses. Visna viruses induce a unique interferon during interaction between lymphocytes and infected macrophages[J]. J Exp Med, 1985, 162(6): 1594-1969.
[17] Steinman R M. The dendritic cell system and its role in immunogenicity[J].Annual Review of Immunology, 1991, 9: 271-296.
[18] Kedzierska K, Mak J, Mijch A, et al. Granulocytemacrophage colonystimulating factor augments phagocytosis of mycobacterium avium complex by human immunodeficiency virus type 1infected monocytes/macrophages in vitro and in vivo[J]. J Infect Dis, 2000, 181(1): 390-394.
[19] Le P T, Lazorick S, Whichard L P, et al. Regulation of cytokine production in the human thymus: epidermal growth factor and transforming growth factor alpha regulate mRNA levels of interleukin 1 alpha(IL 1 alpha), IL 1 beta, and IL 6 in human thymic epithelial cells at a posttranscriptional level[J]. JEM, 1991, 174(5): 1147-1157.
[20] Dai Z, Hooker B S, Quesenberry R D, et al. Optimization of acidothermus cellulolyticus endoglucanase(E1) production in transgenic tobacco plants by transcriptional, post transcription and post translational modification[J]. Transgenic Research, 2005, 14(5), 627-643.
[21] Matsusaka T, Fujikawa K, Nishio Y, et al. Transcription factors NF IL6 and NF kappa B synergistically activate transcription of the inflammatory cytokines, interleukin 6 and interleukin 8[J]. Proc Natl Acad Sci USA, 1993, 90(21): 10193-10197.

备注/Memo

备注/Memo:
收稿日期: 2012-10-08
基金项目: 国家自然科学基金(81272687);科研启动基金(1204807 Y)
作者简介: 韩文正(1986-),男,安徽萧县人,硕士研究生,主要从事肿瘤的靶向基因治疗研究。
通信作者: 周秀梅,电子邮箱:zhouxiumei824@163.com
更新日期/Last Update: