四君子汤及单味药水溶性多糖免疫佐剂活性研究

徐斌; 李妍; 李响; 陈为; 史立言

doi:10.11847/zgggws1112916

四君子汤及单味药水溶性多糖免疫佐剂活性研究

Immunoadjuvent activity of different polysaccharides from Sijunzi decoction and its components

摘要

摘要:
目的探讨四君子汤及4种单味药水溶性多糖对小鼠巨噬细胞增殖、表型及吞噬功能影响。
方法水提醇沉法提取四君子汤水溶性粗多糖，采用滤膜离心法将多糖按分子量（高、中、低）分为3段（分子量分别为 > 100 kDa、30～100 kDa和 < 30 kDa）；相同方法制备组成四君子汤4味单药（人参、白术、茯苓和炙甘草）多糖；用不同多糖（100 mg/L）处理小鼠巨噬细胞（RAW264.7）24 h，流式细胞术分析细胞周期和活性氧（ROS）含量；检测巨噬细胞对荧光微球的吞噬能力；通过流式细胞术检测细胞膜CD40、CD86和CD284（TLR4）表达，分析不同多糖对细胞分化的影响。
结果中、低分子量四君子汤多糖及单味药人参和炙甘草多糖活化巨吞噬细胞的效应较强，可促进细胞增殖，增加细胞S期和G2M期细胞百分率；四君子汤、人参和炙甘草中等分子量多糖组小鼠巨噬细胞CD40阳性细胞百分率分别为、（86.30 ± 3.72）%、（87.19 ± 2.76）%、（85.48 ± 4.63）%明显高于对照组（76.99 ± 3.28）%；CD86阳性细胞百分率分别为（65.60 ± 2.35）%、（65.72 ± 2.35）%、（65.72 ± 2.70）%高于对照组（60.64 ± 2.25）%，而CD284表达量分别为（7.46 ± 0.46）、（6.82 ± 0.16）、（7.54 ± 0.28）高于对照组（6.24 ± 0.74）。
结论四君子汤、人参和炙甘草中等分子量多糖可上调小鼠巨噬细胞CD40、CD86和CD284表达，促进巨噬细胞极化；中、低分子量多糖具有活化巨噬细胞效应。

Abstract:
Objective To study the effect of water-soluble polysaccharides from Sijunzi decoction and its four components on mouse macrophage cells.
Methods The crude polysaccharides from Sijunzi decoction (composed of ginseng, atractylodes, tuckahoe and Radix glycyrrhizae preparata) were extracted with aqueous extraction and alcohol precipitation and then separated into 3 groups of high (> 100 kD), medium (30 to 100 kD), and low (< 30 kD) molecular weight using centrifuge through filter membrane. The same procedures were adopted to acquire crude polysaccharides form the four components of Sijunzi decoction. The mouse macrophage cells RAW264.7 were treated with the extracted polysaccharides at the dosage of 100 mg/L for 24 hours; then the cell cycle, phagocytosis, and reactive oxygen species (ROS) of the cells were determined with flow cytometry (FCM). The expressions of CD40, CD86 and CD284 were also detected with FCM to analyze polarizations of the RAW264.7 cells.
Results The low and medium molecular weight polysaccharides from Sijunzi decoction and its two components (ginseng and Radix glycyrrhizae preparata) promoted the proliferation and increased the proportions of the cells in S and G2M of the RAW264.7 cells. The ratios of CD40 positive cells were 86.30% ± 3.72%, 87.19% ± 2.76%, and 85.48% ± 4.63% for the RAW264.7 cells treated with medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata, and all obviously higher than that (76.99% ± 3.28%) of control cells. Compared to those of the control cells, the RAW264.7 cells treated with medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata also showed higher ratios of CD86 positive (65.60 ± 2.35%, 65.72 ± 2.35%, and 65.72 ± 2.70% vs. 60.64 ± 2.25%) and CD284 positive (7.46 ± 0.46, 7.22 ± 0.14, and 7.54 ± 0.28 vs. 6.24 ± 0.74).
Conclusion The medium molecular weight polysaccharides from Sijunzi decoction, ginseng and Radix glycyrrhizae preparata could upregulate expressions of CD40, CD86 and CD284 and promote the polarizations of RAW264.7 cells; the low and medium molecular weight polysaccharides from Sijunzi decoction and its components have strong immunoadjuvent activity.

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