基于RAGE-TLR4串扰的高糖影响牛肺泡巨噬细胞促炎细胞因子释放的分子机制  

Molecular Mechanism of High Glucose Affecting Proinflammatory Cytokine Release from Bovine Alveolar Macrophages Based on RAGE-TLR4 Crosstalk

作　　者：谭天宇 才冬杰 苟丽萍[1] 任志华[1] 左之才[1] TAN Tianyu;CAI Dongjie;GOU Liping;REN Zhihua;ZUO Zhicai(Sichuan Key Laboratory of Animal Diseases and Human Health, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China)

机构地区：[1]四川农业大学动物医学院,动物疫病与人类健康四川省重点实验室,成都611130

出　　处：《畜牧兽医学报》2021年第10期2944-2952,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省科技计划项目(2018 NZ0002,2019YFQ0012);国家重点研发计划项目(2018YFD0501800);国家现代农业产业技术体系四川肉牛创新团队项目(SCCXTD-2020-13)。

摘　　要：本试验旨在研究高浓度葡萄糖对牛肺泡巨噬细胞(BAMs)促炎细胞因子IL^(-1)β、IL-6及TNF-α释放的影响及其机制是否与RAGE-TLR4相关信号通路串扰有关。将BAMs随机分为正常糖组(NG)、高糖组(HG)、高糖+RAGE抑制剂组(H+F)、高糖+TLR4抑制剂组(H+T)及DMSO组,处理12 h后收集上清及下层细胞。采用qRT-PCR和Western blot检测细胞RAGE、TLR4、MyD88、NF-κB p65的mRNA及蛋白表达情况,ELISA检测上清TNF-α、IL^(-1)β、IL-6浓度。结果表明,高糖极显著上调RAGE、TLR4、MyD88和NF-κB p65基因、蛋白表达水平以及上清液中IL^(-1)β、IL-6、TNF-α浓度(P<0.01);RAGE抑制剂与TLR4抑制剂均极显著抑制高糖引起的RAGE、TLR4、MyD88和NF-κB p65基因、蛋白表达水平上调以及IL^(-1)β、IL-6、TNF-α释放(P<0.01),即RAGE与TLR4均在激活RAGE/TLR4/MyD88/NF-κB炎症信号通路中发挥调控作用。综上所述,高糖能够通过RAGE-TLR4串扰引起牛肺泡巨噬细胞释放促炎细胞因子IL^(-1)β、IL-6及TNF-α,进一步阐明了高糖促进牛肺泡巨噬细胞炎症反应的分子机制。In this study,we aimed to investigate whether high glucose regulates proinflammatory cytokine IL^(-1)β,IL-6 and TNF-αrelease from bovine alveolar macrophages(BAMs)through RAGE-TLR4 crosstalk.BAMs were randomly divided into normal glucose group(NG),high glucose group(HG),high glucose+RAGE inhibitor group(H+F),high glucose+TLR4 inhibitor group(H+T)and DMSO group,and the supernatant and lower cells were collected after 12 h of treatment.The mRNA and protein expression of RAGE,TLR4,MyD88 and NF-κB p65 were detected by qRT-PCR and Western blot,and the concentrations of TNF-α,IL^(-1)βand IL-6 in the supernatant were detected by ELISA.The levels of mRNA and protein expression of RAGE,TLR4,MyD88 and NF-κB p65 and the concentrations of IL^(-1)β,IL-6 and TNF-αin the supernatant of HG group were significantly higher than those of NG group,H+F group and H+T group(P<0.01);the levels of mRNA and protein of MyD88,NF-κB p65 and RAGE and the concentrations of three proinflammatory cytokines in the supernatant of H+F group and H+T group were significantly higher than those of NG group(P<0.05,P<0.01).Our results suggest that high glucose can cause the release of proinflammatory cytokines IL^(-1)β,IL-6 and TNF-αfrom bovine alveolar macrophages via RAGE-TLR4 crosstalk,which further elucidated the molecular mechanism of high glucose promoted the inflammatory response in bovine alveolar macrophages.

关 键 词：炎症信号通路 RAGE-TLR4串扰 牛肺泡巨噬细胞 高糖 

分 类 号：S852.33[农业科学—基础兽医学]