期刊文献+

大鼠骨髓树突状细胞的分离培养及HMGB1/TLR4信号通路的调控作用 预览

Study on isolation and culture of rat bone marrow-derived dendritic cells: regulatory effect of HMGB1/TLR4 signaling pathway
在线阅读 下载PDF
分享 导出
摘要 目的:研究HMGB1/TLR4信号通路对大鼠骨髓树突状细胞(mDCs)MyD88、NF-κBP65、细胞因子、共刺激分子表达的影响,探讨此信号通路是否对m DCs具有调控作用。方法:mDCs分离、培养、纯化、鉴定后,随机分为5组,每组各设24、48、72h3个时间点。C组:m DCs加于RPMIl640完全培养基中;H组:加入HMGB1;H-Ab组:加入HMGB1后30min再加入HMGB1特异性中和抗体;TLR4-A组:加入HMGB1后30min再加入TLR4拮抗剂;IgG组:加入HMGB1后30min再加入对照IgG抗体。检测各组各时间点m DCsMy D88、NF-κBP65蛋白和基因、共刺激分子(CD80、CD86)的表达及上清液中细胞因子(IL-6、IL-8、IL-12、TNF-α)的浓度变化。结果:24h、48h、72h各组HMGB1刺激后MyD88、NF-κBP65蛋白和基因、细胞因子、共刺激分子表达明显升高,使用HMGB1特异性中和抗体、TLR4拮抗剂后明显下降,差异均有统计学意义(P〈0.01)。结论:HMGB1通过m DCs的受体TLR4影响下游MyD88、NF-κBP65蛋白和基因的表达,并刺激细胞因子的分泌,同时促进共刺激分子CD80、CD86的表达,对mDCs起一定的调控作用。 Objective: To investigate the regulatory effect of HMGB1/TLR4 signaling pathway on the expression of My D88, NF-κB P65, cytokine and costimulatory molecules in rat bone marrow-derived dendritic cells(m DCs). Methods: Dendritic cells were isolated, cultured, purified and identified in vitro from bone marrow stem cells of rats, then randomly divided into 5 groups: m DCs cultured with HMGB1 only(H group), HMGB1 specific neutralizing antibody(H-Ab group), TLR4 antagonist(TLR4-A group) and control Ig G(Ig G group) 30 min after HMGB1, separately, or with RPMI l640 serum-containing medium as control group(C gruop). Each group was observed at different time points of 24 h, 48 h and 72 h by detecting the expression of My D88, NF-κB P65 protein and m RNA, costimulatory molecules(CD80, CD86) and the levels of cytokine(IL-6, IL-8, IL-12, TNF-α) in the supernatant. Results: My D88 and NF-κB P65 protein and m RNA, costimulatory molecules and cytokine levels in m DCs stimulated with HMGB1 were significantly higher than control group at each time point(P〈0.01). The levels were significantly decreased after being treated with HMGB1 specific neutralizing antibody or TLR4 antagonist(P〈0.01). Conclusion: Data analysis suggests that HMGB1 plays a regulatory role in m DCs by affecting the downstream My D88, NF-κB P65 protein and m RNA expression through TLR4, stimulating the secretion of cytokine and promoting the expression of costimulatory molecules(CD80, CD86).
作者 王涵磊 薛继阳 葛捍伟 夏杰 林炜 赵琦峰 WANG Hanlei, XUE Jiyang, GE Hanwei, XIA Jie, LIN Wei, ZHAO Qifeng. (Department of Cardiovascular and Thoracic Surgery, Pediatric Heart Center, the Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, 325027)
出处 《温州医科大学学报》 CAS 2018年第7期508-516,共9页 Journal of Wenzhou Medical College
基金 浙江省科技厅实验动物科技计划项目(2015C37133)
关键词 高迁移率族蛋白B1 TOLL样受体4 信号通路 树突状细胞 大鼠 high mobility group box 1 protein Toll-like receptor-4 signaling pathway dendritic cells rats
作者简介 王涵磊(1993-),男,浙江温州人,硕士生。;通信作者:赵琦峰,主任医师,Email:zhaoqf1862@163.com。
  • 相关文献
投稿分析

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部 意见反馈