子宫珠蛋白结合蛋白在antiflammin-1促进LPS诱导的RAW264.7细胞IL-10表达和分泌中的作用

Role of uteroglobin-binding protein in antiflammin-1 promoting IL-10 expression and secretion in RAW264.7 cells induced by endotoxin

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 present study investigated the effect of antiflammin-1 （AF-1） on LPS-induced IL-10 secretion from RAW264.7 cells through uteroglobin-binding protein （UGBP）. Cultured RAW264.7 cells, a murine monocyte-macrophage cell line, were divided as following： control group, LPS group （1μg/mL LPS）, AF-1 group （100 μmol/L AF-1）, LPS＋AF-1 group （2 h of 100 μmol/L AF-1 pre- treatment before LPS addition）, and LPS＋AF-I＋anti-UGBP group （30 min of anti-UGBP antibody pretreatment before successive treatments with AF-1 and LPS）. IL-10 concentration in the supernatants was detected by ELI SA assay, and the level of IL-10 mRNA expression in macrophage was detected by using RT-PCR method. The results showed that AF-1 significantly increased LPS-induced IL-10 secretion in RAW264.7 cells in a dose dependent way, and up-regulated its mRNA level. Anti-UGBP antibody pretreatment attenuated the augmented effect of AF-1 on LPS-induced IL-10 secretion and gene expression. These results suggest that AF-1 promotes LPS-induced IL-10 secretion from macrophages, and this effect is mediated by UGBP.