青蒿琥酯增加小鼠腹腔巨噬细胞内化内毒素、大肠埃希菌的作用及可能机制

中国临床药理学与治疗学 ›› 2012, Vol. 17 ›› Issue (1): 40-46.

青蒿琥酯增加小鼠腹腔巨噬细胞内化内毒素、大肠埃希菌的作用及可能机制

余梦辰, 潘夕春, 李小丽, 李军, 肖康康, 李斌, 周红

第三军医大学药学院药理学教研室,400038 重庆

收稿日期:2011-11-11 修回日期:2011-12-10 出版日期:2012-01-26 发布日期:2012-02-16
通讯作者: 李斌,男,博士,副教授,研究方向:感染、炎症的发病机制及其防治措施。E-mail: libin6033@yahoo.com.cn;周红,女,博士,教授,博士研究生导师,研究方向:感染、炎症的发病机制及其防治措施。Tel: 023-68752266 E-mail: zhouh64@163.com
作者简介:余梦辰,女,在读硕士研究生,研究方向:感染、炎症的发病机制及其防治措施。Tel: 023-68752265 E-mail: agnes15@163.com
基金资助:
国家自然科学基金青年科学基金项目(81001440)

Artesunate enhance the ability of peritoneal macrophages endocytosing LPS and Escherichia coli and its mechnisms

YU Meng-chen, PAN Xi-chun, LI Xiao-li, LI Jun, XIAO Kang-kang, LI Bin, ZHOU Hong

Department of Pharmacology, College of Pharmacy, Third Military Medical University, Chongqing 400038, China

Received:2011-11-11 Revised:2011-12-10 Online:2012-01-26 Published:2012-02-16

摘要/Abstract

摘要： 目的: 观察青蒿琥酯(artesunate, AS)对小鼠腹腔巨噬细胞内化清除脂多糖/内毒素(lipopolysaccharide/endotoxin, LPS)和吞噬大肠埃希菌的影响,并初步探讨其可能的作用机制。方法: MTT法检测不同浓度网格蛋白抑制剂(monodansylcadaverine, MDC)、内体酸化抑制剂氯喹(chloroquine, CQ)对小鼠腹腔巨噬细胞活力的影响,以选择恰当的药物工作浓度;激光共聚焦法检测青蒿琥酯及MDC、CQ对小鼠腹腔巨噬细胞内化FITC-LPS的影响;分别采用激光共聚焦和菌落集落形成计数实验观察青蒿琥酯对小鼠腹腔巨噬细胞内化大肠埃希菌的影响;逆转录PCR检测青蒿琥酯对小鼠腹腔巨噬细胞清道夫受体A(class A scavenger receptor,SR-A) mRNA表达的影响。结果: MDC和CQ浓度分别小于 25 μg/mL 和 20 μg/mL 时对小鼠腹腔巨噬细胞活力无影响;MDC、CQ可抑制小鼠腹腔巨噬细胞内化LPS,青蒿琥酯可增加小鼠腹腔巨噬细胞对LPS的内化,而且青蒿琥酯可增加MDC和CQ处理的巨噬细胞内化LPS的功能。激光共聚焦和菌落集落形成计数实验均显示青蒿琥酯可增加小鼠腹腔巨噬细胞对大肠埃希菌的内化能力。逆转录PCR结果显示青蒿琥酯可增强LPS处理或未处理的小鼠腹腔巨噬细胞SR-A mRNA的表达。结论: 青蒿琥酯可增强小鼠腹腔巨噬细胞内化LPS、大肠埃希菌的能力,该作用可能与SR-A mRNA表达升高有关。

关键词: 青蒿琥酯, 内化, 脂多糖, 大肠埃希菌, 清道夫受体

Abstract: AIM: To investigate the effect of artesunate (AS) on peritoneal macrophages endocytosing lipopolysaccharide/endotoxin (LPS) and phagocytosing Escherichia coli.METHODS: Effects of various concentrations of monodansylcadaverine (MDC) and chloroquine(CQ) on the viability of peritoneal macrophages were observed by MTT test. Effects of artesunate, monodansylcadaverine and chloroquine on LPS internalization of peritoneal macrophages were detected by confocal laser scaning microscopy. The effect of artesunate on Escherichia coli internalization of peritoneal macrophages was examined by confocal laser scaning microscopy and bacterial clone forming counting test. The effect of artesunate on class A scavenger receptor (SR-A) mRNA expression was measured using RT-PCR method.RESULTS: 25 μg /mL of MDC and 20 μg /mL of chloroquine had no effect on the viability of peritoneal macrophages. Artesunate could increase LPS internalization of peritoneal macrophages, while MDC and chloroquine could inhibit LPS but artesunate could still increase LPS internalization of macrophages treated with MDC and chloroquine. Artesunate could increase Escherichia coli internalization, and artesunate could up-regulate SR-A mRNA expression of peritoneal macrophages treated with or without LPS.CONCLUSION: Artesunate could enhance the ability of the peritoneal macrophages to internalize LPS and Escherichia coli, which was associated with up-regulation of SR-A mRNA expression of the peritoneal macrophages.

Key words: Artesunate, Internalization, Lipopolysaccharide, Escherichia coli, Scavenger receptor

中图分类号:

R965.2

余梦辰, 潘夕春, 李小丽, 李军, 肖康康, 李斌, 周红. 青蒿琥酯增加小鼠腹腔巨噬细胞内化内毒素、大肠埃希菌的作用及可能机制[J]. 中国临床药理学与治疗学, 2012, 17(1): 40-46.

YU Meng-chen, PAN Xi-chun, LI Xiao-li, LI Jun, XIAO Kang-kang, LI Bin, ZHOU Hong. Artesunate enhance the ability of peritoneal macrophages endocytosing LPS and Escherichia coli and its mechnisms[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2012, 17(1): 40-46.

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