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本文旨在观察antiflammin-1(AF-1)预处理对脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7细胞白细胞介素10(interleukin-10,IL-10)分泌的影响,并探讨子宫珠蛋白结合蛋白(uteroglobin-binding protein,UGBP)在介导AF-1对LPS诱导的巨噬细胞IL-10分泌及表达影响中的作用。将RAW264.7细胞分为空白对照组、LPS组(1μg/mL LPS)、AF-1组(100μmol/LAF-1)、AF-1+LPS组(100μmol/LAF-1预处理2h后,加入LPS)及抗UGBP抗体+AF-1+LPS组(抗-UGBP抗体预处理30min后,再依次用AF-1和LPS处理),用ELISA法、RT-PCR法分别检测各组IL-10蛋白分泌和mRNA表达。结果显示,AF-1预处理呈剂量依赖性显著增加LPS诱导的RAW264.7细胞IL-10分泌,并上调IL-10mRNA水平;而抗UGBP抗体预处理可显著抑制AF-1的促IL-10分泌和上调mRNA表达作用。以上结果提示,AF-1可促进LPS诱导的巨噬细胞IL-10的分泌和mRNA表达,且该作用依赖于UGBP的介导。
The aim of this study was to investigate the effect of antiflammin-1 pretreatment on the secretion of interleukin-10 (IL-10) in RAW264.7 cells induced by lipopolysaccharide (LPS) UGBP plays an important role in mediating the effect of AF-1 on IL-10 secretion and its expression induced by LPS in macrophages. RAW264.7 cells were divided into blank control group, LPS group (1μg / mL LPS), AF-1 group (100μmol / LAF-1) and AF-1 + LPS group LPS) and anti-UGBP antibody + AF-1 + LPS group (anti-UGBP antibody pretreatment 30min, followed by AF-1 and LPS followed by treatment), ELISA, RT- Secretion and mRNA expression. The results showed that pretreatment with AF-1 significantly increased IL-10 secretion and IL-10 mRNA level induced by LPS in RAW264.7 cells in a dose-dependent manner. Anti-UGBP pretreatment significantly inhibited the production of IL-10 Secretion and upregulation of mRNA expression. The above results suggest that AF-1 can promote LPS-induced IL-10 secretion and mRNA expression in macrophages, and this effect is dependent on the mediation of UGBP.