17β-雌二醇抑制LPS诱导Raw264.7细胞炎症反应及其作用机制  

Inhibitory effect of 17β-estradiol on LPS-induced inflammation in Raw264.7 cells and its regulatory mechanism

作　　者：余紫薇 王晓杜 罗通旺 姜胜 巴少波 王磊 宋泉江 周彬 宋厚辉 邵春艳 YU Ziwei;WANG Xiaodu;LUO Tongwang;JIANG Sheng;BA Shaobo;WANG Lei;SONG Quanjiang;ZHOU Bin;SONG Houhui;SHAO Chunyan(Key Laboratory of Applied Technology on Green-Eco-Healthy Animal Husbandry of Zhejiang Province/Zhejiang Provincial Engineering Research Center for Animal Health Diagnostics&Advanced Technology/Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management/China-Australia Joint Laboratory for Animal Health Big Data Analytics,College of Animal Science and Technology/College of Veterinary Medicine,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学动物科技学院/动物医学院浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程研究中心/浙江省动物医学与健康管理国际科技合作基地/中澳动物健康大数据分析联合实验室,浙江杭州311300

出　　处：《中国兽医学报》2022年第4期756-762,769,共8页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金青年基金资助项目(31802258);浙江农林大学科研发展基金资助项目(2018FR015,2015FR042,2020FR045);浙江省自然科学基金资助项目(LQ20C180002);浙江省教育厅科研基金资助项目(Y202044917)。

摘　　要：为探究雌二醇(17β-E2)对脂多糖(LPS)诱导的小鼠Raw264.7细胞炎症反应的影响及作用机制,使用LPS刺激Raw264.7诱导细胞炎症为模型,试验分为Mock、LPS、E2和E2+LPS组,分别用qRT-PCR方法检测各组TLR4、IL-1β、IL-18和NLRP3 mRNA表达水平,用ELISA法检测各组细胞上清中IL-1β分泌量变化,用Western blot法检测各组细胞中IL-1β、NF-κB p65、p-IκBα和IκBα的蛋白表达水平,用免疫荧光法观察NF-κB p65在细胞内的表达和定位。结果显示,17β-E2可以不同程度的抑制LPS诱导的Raw264.7细胞TLR4、IL-1β、IL-18和NLRP3 mRNA的表达,差异显著(P<0.05),抑制细胞内IL-1β的蛋白表达量和细胞上清中IL-1β的分泌量,差异极显著(P<0.01);Western blot结果显示17β-E2可以降低LPS诱导的IκBα的磷酸化水平,差异极显著(P<0.01),抑制NF-κB p65进入细胞核;免疫荧光结果显示17β-E2主要通过ERβ抑制p65入核。综上所述,17β-E2主要通过ERβ抑制NF-κB p65入核,从而发挥抑制LPS诱导的Raw264.7细胞炎症反应的作用。The purpose of this study was to explore the effect and mechanism of 17β-estradiol(17β-E2)on the inflammatory response of Raw264.7 cells induced by lipopolysaccharide(LPS).Raw264.7 cells inflammation models induced with LPS were used in this study.Raw264.7 cells were divided into control group,LPS group,E2 group,E2+LPS group,the mRNA expression levels of TLR4,IL-1β,IL-18 and NLRP3 were detected by qRT-PCR.The protein levels of IL-1β,NF-κB p65,p-IκBαand IκBαwere measured by Western blot.The content of IL-1βin the cell supernatant were measured by ELISA.The expression and localization of NF-κB p65 in cells were observed using immunofluorescence.The results showed that 17β-E2 significantly decreased the expression of LPS induced TLR4,IL-1β,IL-18 and NLRP3 mRNA level(P<0.05);17β-E2 significantly decreased LPS induced the protein levels of IL-1βin cells and supernatant(P<0.01).Moreover,the decreased level of phosphorylation IκBαand the declined translocation into nucleus of NF-κB p65were observed in E2+LPS group cells.In conclusion,17β-E2 showed an inhibitory effect on LPS induced inflammation in Raw264.7cells by inactivation NF-κB p65mainly through ERβ.

关 键 词：17Β-E2 LPS RAW264.7 细胞炎症 NF-κB信号通路 

分 类 号：S859.79[农业科学—临床兽医学]