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目的探讨红车轴草提取物(Trifoliumpratense Leguminosae extract,TLE)体外对小鼠淋巴细胞[Ca2+]i及腹腔巨噬细胞NO分泌和吞噬微球的影响。方法无菌条件下制备小鼠淋巴细胞悬液及小鼠腹腔巨噬细胞悬液;MTT法检测药物对细胞悬液的毒性情况;Fluo-4/AM染色结合流式细胞术分析TLE对小鼠淋巴细胞[Ca2+]i的影响;Griess反应系统检测TLE对脂多糖(LPS)刺激的小鼠腹腔巨噬细胞NO分泌的影响;1μm与2μm直径的荧光微球结合流式细胞术分析TLE对小鼠腹腔巨噬细胞吞噬作用的影响。结果终质量浓度为20、40mg/L的TLE对细胞的毒性小;TLE促进了淋巴细胞的Ca2+内流;TLE抑制了巨噬细胞的NO分泌与吞噬作用,与非TLE组比较P<0.01。结论对淋巴细胞[Ca2+]i及巨噬细胞的NO分泌和吞噬的作用可能是TLE调节小鼠免疫系统的途径。
Objective To investigate the effects of Trifoliumpratense Leguminosae extract (TLE) on NO production and phagocytosis of lymphocytes [Ca2+]i and peritoneal macrophages in mice. METHODS: Mouse lymphocyte suspensions and mouse peritoneal macrophage suspensions were prepared under aseptic conditions; MTT assay was used to detect the toxicity of the drugs to cell suspensions; Fluo-4/AM staining was combined with flow cytometry to analyze TLEs against mice. Effects of lymphocyte [Ca2+]i; Griess reaction system to detect the effect of TLE on lipopolysaccharide (LPS)-stimulated NO secretion in mouse peritoneal macrophages; 1μm and 2μm diameter fluorescent microspheres combined with flow cytometry analysis of TLE vs. small Effect of phagocytosis of murine peritoneal macrophages. Results TLE with a final concentration of 20 and 40 mg/L was less toxic to cells; TLE promoted Ca2+ influx of lymphocytes; TLE inhibited the secretion and phagocytosis of NO in macrophages, which was compared with non-TLE group P<0.01. Conclusions The effects of NO secretion and phagocytosis on lymphocytic [Ca2+]i and macrophages may be the pathway by which TLE regulates the immune system in mice.