中文摘要：

探讨采用高内涵筛选技术检测巨噬细胞（RAW264.7）吞噬荧光标记大肠杆菌（GFP-Escherichia coli）方法的灵敏性、稳定性及可靠性,建立中药促进巨噬细胞吞噬活性的高内涵评价方法。对比考察不同的感染复数（multiplicity of infection,MOI）即细菌和细胞比值,以及不同孵育时间对巨噬细胞吞噬细菌能力的影响（吞噬指数=被吞噬的细菌荧光强度/巨噬细胞数）,优化高内涵筛选方法和实验参数,并用于测定铜皮石斛对巨噬细胞吞噬作用的影响。结果表明,吞噬指数与MOI值呈正相关,孵育时间在1.5 h吞噬指数最高,优化后的条件：细胞数1×10~4个/孔,孵育时间1.5 h,MOI值为50（即细菌数为5×10~5个/孔）,该条件下检测结果的RSD〈10%。采用所建立的方法检测铜皮石斛调节巨噬细胞吞噬作用实验的结果显示,在0.31~2.50 g·L~（-1）内铜皮石斛水提液对巨噬细胞吞噬具有剂量依赖的促进作用。本文建立了巨噬细胞吞噬作用评价的高内涵筛选方法,并进一步证明了铜皮石斛水提物促进巨噬细胞的吞噬作用。

英文摘要：

The aim of the present study was to explore a sensitive, stable and reliable method for evaluating the phagocytosis, in which RAW264.7 macrophages engulfed GFP-Escherichia coli was tested by high-content screening technology. The study was conducted to optimize the method in evaluation of traditional Chinese medicine in the promotion of macrophage function. By testing macrophages at different ratio of bacteria to cells （multiplicity of infection, MOI）, and at different incubation time, we optimized a high content screening method and the experimental parameters to determine the impact of bacteria in macrophages （fluorescence intensity index =be swallowed bacteria/macrophages）. The method was used to determine whether Dendrobium moniliforme （DM） have effects on macrophage phagocytosis. The results show that the index has a positiverelationship with MOI values, and the highest index was observed at incubation time of 1.5 h. The optimized conditions was 1 × 10^4 cells/well with a MOI of 50 ： 1 （bacteria ： cells） with incubation of 1.5 h. Under this condition, the relative standard deviation （RSD） was less than 10% in the precision test. Using the method to detect DM regulating macrophage phagocytosis experiment results showed that in 0.31-2.50 g.L^-1 concentration range, DM has a dose-response effect in promoting phagocytosis. We successfully established the method for evaluation of macrophage phagocytosis, and proved the activity of DM in promotion of macrophage phagocytosis.