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目的建立巨噬细胞甘露糖受体(MMR)结合物筛选模型,用于筛选以甘露糖受体(MR)为靶标的潜在活性物质。方法将小鼠腹腔巨噬细胞分别与不同浓度的D-甘露糖和D-半乳糖共孵,用流式细胞仪和荧光显微镜检测D-甘露糖和D-半乳糖对Mφ结合异硫氰酸荧光素标记的甘露糖化牛血清白蛋白(M-FITC-BSA)的拮抗作用,优化实验条件,建立MMR筛选模型。结果两种检测方法均显示:随D-甘露糖浓度的上升,M-FITC-BSA标记的Mφ检出率逐步下降(P<0.01),而这种现象在D-半乳糖组中未观察到;当D-甘露糖达到0.080mmol.L-1浓度时,便能明显拮抗Mφ结合M-FITC-BSA(P<0.05或P<0.01)。中药多糖的筛选结果显示:在一定浓度下,黄芪、白术、防风、大枣等中药多糖组分能浓度依赖性地拮抗小鼠腹腔Mφ与M-FITC-BSA的结合(P<0.05或P<0.01)。结论MMR模型对MR结合成分的筛选作用稳定而敏感,对快速筛选中药免疫调节成分、研究中药免疫调节机制具有重要意义。
Objective To establish a macrophage mannose receptor (MMR) conjugate screening model for the screening of potential active substances targeting the mannose receptor (MR). Methods Mouse peritoneal macrophages were co-incubated with different concentrations of D-mannose and D-galactose, and D-mannose and D-galactose were detected by flow cytometry and fluorescence microscopy. The antagonism of fluorescein-labeled mannosylated bovine serum albumin (M-FITC-BSA) was optimized and experimental conditions were established to establish an MMR screening model. Results Both methods showed that the detection rate of M-FITC-BSA-labeled Mφ decreased gradually with increasing D-mannose concentration (P<0.01), but this phenomenon was not observed in the D-galactose group. When D-mannose reached a concentration of 0.080 mmol.L-1, it significantly antagonized Mφ binding to M-FITC-BSA (P<0.05 or P<0.01). The screening results of polysaccharides from traditional Chinese medicines showed that polysaccharides of Radix Astragali, Atractylodes macrocephala, windbreak, and jujube could inhibit the binding of Mφ and M-FITC-BSA in a dose-dependent manner (P<0.05 or P<0.01). 0.01). Conclusion The MMR model has a stable and sensitive screening effect on MR binding components. It is of great significance for the rapid screening of immunomodulatory components of traditional Chinese medicine and the study of immunoregulatory mechanisms of traditional Chinese medicine.