粗根荨麻多糖对巨噬细胞吞噬功能和极化的影响

伍根瑞; 马建成; 李继兴; 罗娅; 包爽; 王重娟; 吴明一

doi:10.13386/j.issn1002-0306.2023110037

粗根荨麻多糖对巨噬细胞吞噬功能和极化的影响

Effects of Polysaccharides from Urtica macrorrhiza Hand.-Mazz. on Phagocytic Function and Polarization of Macrophage

摘要

摘要: 目的：研究粗根荨麻鼠李半乳糖醛酸聚糖（Urtica macrorrhiza Hand.-Mazz. Rhamnnogalacturonans，UMRG）和粗根荨麻阿拉伯半乳聚糖（Urtica macrorrhiza Hand.-Mazz. Arabinogalactan，UMAG）对巨噬细胞的吞噬功能和极化的影响。方法：CCK8法研究粗根荨麻多糖UMRG、UMAG对巨噬细胞活力的影响；中性红实验、流式细胞术、激光共聚焦研究UMRG、UMAG对巨噬细胞吞噬功能的影响；流式细胞术研究UMRG、UMAG对巨噬细胞极化的影响；Griess法和酶联免疫吸附法（Enzyme-linked immunosorbent assay，ELISA）检测UMRG、UMAG对巨噬细胞一氧化氮（Nitric oxide，NO）、肿瘤坏死因子-α（Tumor necrosis factor-α，TNF-α）、白细胞介素6（Interleukin 6，IL-6）、白细胞介素1β（Interleukin 1β，IL-1β）分泌水平的影响。结果： UMRG和UMAG在10~320 µg/mL浓度范围内对巨噬细胞均无显著的细胞毒性；UMRG和UMAG均可显著增强巨噬细胞对中性红和模式抗原的吞噬作用（P<0.05），其中UMRG促进巨噬细胞吞噬的作用较UMAG强。UMRG和UMAG均能极显著刺激巨噬细胞CD86和CD206分子表达量的上调（P<0.01），其中CD86上调比例显著高于CD206，UMRG的作用较UMAG强；UMRG、UMAG均能极显著刺激巨噬细胞分泌NO、TNF-α、IL-6（P<0.01）。结论：粗根荨麻多糖UMRG、UMAG可激活巨噬细胞，增强巨噬细胞的吞噬功能，主要促进巨噬细胞向M1型极化，增强免疫活性，且UMRG活性强于UMAG。

Abstract: Objective: To investigate the effects of Urtica macrorrhiza Hand.-Mazz. rhamnnogalacturonans (UMRG), and Urtica macrorrhiza Hand.-Mazz. arabinogalactan (UMAG) on the immunological function by regulating phagocytic and polarization of macrophages. Methods: The effects of UMRG and UMAG on the viability of macrophage were studied by the cell counting kit-8 (CCK-8). The effects of UMRG and UMAG on the phagocytosis of macrophage were investigated by neutral red assay, flow cytometry and laser confocal microscopy. The effects of UMRG and UMAG on macrophage polarization were studied by flow cytometry. The effects of UMRG and UMAG on the secretion of nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) were detected by Griess assay and enzyme-linked immunosorbent assay (ELISA). Results: UMRG and UMAG exhibited no significant cytotoxicity to macrophages in the concentration range of 10~320 µg/mL. Both UMRG and UMAG significantly enhanced the macrophage phagocytosis of neutral red and model antigens (P<0.05) by macrophage, and the activity of UMRG was stronger than UMAG. Both UMRG and UMAG significantly up-regulated the expression of CD86 and CD206 molecules on macrophage (P<0.01), however, the up-regulation proportion of CD86 was significantly higher than that of CD206, and the activity of UMRG was stronger than UMAG. Both UMRG and UMAG stimulated the secretion of NO, TNF-α, and IL-6 by macrophage (P<0.01). Conclusion: UMRG and UMAG from Urtica macrorrhiza Hand.-Mazz. can activate macrophage, enhance its phagocytic function and predominantly promote its polarization towards M1 type, thus they may be natural imunopotentiators. The activity of UMRG is stronger than UMAG.