| 摘要: |
| 目的:评价黄芪甲苷(AS-Ⅳ)对脂多糖(LPS)调节巨噬细胞M1/M2型极化的影响及其相关机制。方法:体外培养小鼠单核巨噬细胞系RAW264.7，采用随机数字表法分为3组:对照组(C组)、LPS组、AS-Ⅳ干预(AS-Ⅳ)组。C组细胞正常培养，LPS组加入终浓度为100 ng/mL的LPS处理24 h，AS-Ⅳ组加入终浓度为100 ng/mL的LPS和200 μg/mL的AS-Ⅳ处理24 h。利用ELISA法检测白细胞介素(IL)-1β、IL-10和IL-18水平，qRT-PCR法检测CD86、CD11c、CD206和精氨酸酶-1(Arg-1)的mRNA表达，流式细胞术检测CD86(+)和CD206(+)细胞百分比，Western blot法检测NOD样受体热蛋白结构域相关蛋白3(NLRP3)、凋亡相关斑点样蛋白(ASC)和半胱氨酸蛋白酶-1(caspase-1)的蛋白表达。结果:与C组比较，LPS组IL-1β和IL-18水平、CD86和CD11c mRNA表达、CD86(+)细胞百分比、NLRP3、ASC和caspase-1蛋白表达升高，IL-10水平、CD206和Arg-1 mRNA表达、CD206(+)细胞百分比降低(P ＜0.05)；与LPS组比较，AS-Ⅳ组IL-1β和IL-18水平、CD86和CD11c mRNA表达、CD86(+)细胞百分比、NLRP3、ASC和caspase-1蛋白表达降低，IL-10水平、CD206和Arg-1 mRNA表达、CD206(+)细胞百分比升高(P ＜0.05)。结论:AS-Ⅳ能够抑制LPS诱导的巨噬细胞M1型极化并促进其向M2型极化，其机制与下调NLRP3炎症小体表达有关。 |
| 关键词: 黄芪甲苷 脂多糖 巨噬细胞 炎症 脓毒症 |
| DOI：10.3969/j.issn.1007-6948.2024.06.026 |
| 投稿时间：2024-04-11 |
| 基金项目:国家自然科学基金青年项目(82102248) |
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| Effect of Astragaloside Ⅳ on lipopolysaccharide-induced macrophages M1/M2 polarization and related mechanism |
| LU Fu-tai,YANG Tao,LI Man |
| Department of Anesthesiology, Tianjin Union Medical Center, Tianjin300121, China |
| Abstract: |
| Objective To evaluate the effect of Astragaloside Ⅳ (AS-Ⅳ) on lipopolysaccharide (LPS)-induced macrophages M1/M2 polarization and related mechanism. Methods Mouse monocyte-derived macrophages RAW264.7 were cultured in vitro and were randomly divided into 3 groups: control group (group C), LPS group (group LPS) and AS-Ⅳ treatment group (group AS-Ⅳ). In group C, cells were cultured in the common culture medium. In group LPS, 100 ng/mL LPS was added into the culture medium for 24 h incubation. In group AS-Ⅳ, 100 ng/mL LPS and 200 μg/mL AS-Ⅳ were added into the culture medium for 24 h incubation. The levels of IL-1β, IL-10 and IL-18 in culture medium were measured by ELISA assay. The mRNA levels of CD86, CD11c, CD206 and Arginase-1(Arg-1) were measured by qRT-PCR assay. The percentage of CD86(+) and CD206(+) cells was assessed by flow cytometry. The expression levels of nucleotide-binding oligomerization domain-like receptor containing pyrin domain 3 (NLRP3) and apoptosis-associated speck-like protein (ASC) were detected by Western blot assay. Results Compared with group C, the levels of IL-1β and IL-18, the mRNA levels of CD86 and CD11c, the percentage of CD86(+) cells and the expression levels of NLRP3, ASC and caspase-1 were increased, while the level of IL-10 and the mRNA levels of CD206 and Arg-1, and the percentage of CD206(+) cells were decreased in group LPS(P <0.05). Compared with group LPS, the levels of IL-1β and IL-18, the mRNA levels of CD86 and CD11c, the percentage of CD86(+) cells and the expression levels of NLRP3, ASC and caspase-1 were decreased, while the level of IL-10 and the mRNA levels of CD206 and Arg-1, and the percentage of CD206(+) cells were increased in group AS-Ⅳ(P <0.05). Conclusion AS-Ⅳ can inhibit LPS-induced macrophages M1 polarizaion and promote macrophages M2 polarizaion, whose mechanism may be associated with down-regulating NLRP3 inflammasome expression. |
| Key words: Astragaloside Ⅳ lipopolysaccharide macrophage inflammation sepsis |
