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

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

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(li-popolysaccharide/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)mR-NA表达的影响。结果:MDC和CQ浓度分别小于25μg/mL和20μg/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。

ABSTRACT AIM： To investigate the effect of artesunate （AS） on peritoneal macrophages en- docytosing lipopolysaccharide/endotoxin （LPS） and phagocytosing Escherichia coli. METHODS： Effects of various concentrations of monodansyl- cadaverine （MDC） and chloroquine（CQ） on the viability of peritoneal macrophages were ob- served 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 in- ternalization of peritoneal macrophages was ex- amined 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 RTPCR method. RESULTS：25 μg/mL of MDC and 20 μg /mL of chloroquine had no effect on the viability of peritoneal macrophages. Artesunatecould 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 in- crease Escherichia coli internalization, and arte- sunate 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 upregulation of SR-A mRNA expression of the peritoneal macrophages.