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

doi:10.11843/j.issn.0366-6964.2021.010.025

畜牧兽医学报 ›› 2021, Vol. 52 ›› Issue (10): 2944-2952.doi: 10.11843/j.issn.0366-6964.2021.010.025

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

谭天宇, 才冬杰, 苟丽萍, 任志华, 左之才*

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

收稿日期:2021-01-18 出版日期:2021-10-23 发布日期:2021-10-27
通讯作者: 左之才,主要从事动物营养代谢性疾病研究,E-mail:zzcjl@126.com
作者简介:谭天宇(1996-),男,四川广安人,硕士生,主要从事中西兽医与临床研究,E-mail:13350907018@163.com
基金资助:
四川省科技计划项目（2018NZ0002；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）

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

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

Received:2021-01-18 Online:2021-10-23 Published:2021-10-27

摘要/Abstract

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-1β、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-1β、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-1β、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-1β、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-1β、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。

关键词: 炎症信号通路, RAGE-TLR4串扰, 牛肺泡巨噬细胞, 高糖

Abstract: In this study, we aimed to investigate whether high glucose regulates proinflammatory cytokine IL-1β, IL-6 and TNF-α release 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-1β and 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-1β, IL-6 and TNF-α in 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-1β, IL-6 and TNF-α from bovine alveolar macrophages via RAGE-TLR4 crosstalk, which further elucidated the molecular mechanism of high glucose promoted the inflammatory response in bovine alveolar macrophages.

Key words: inflammatory signaling pathway, RAGE-TLR4 crosstalk, bovine alveolar macrophage, high glucose

中图分类号:

S852.33

谭天宇, 才冬杰, 苟丽萍, 任志华, 左之才. 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[J]. 畜牧兽医学报, 2021, 52(10): 2944-2952.

TAN Tianyu, CAI Dongjie, GOU Liping, REN Zhihua, ZUO Zhicai. Molecular Mechanism of High Glucose Affecting Proinflammatory Cytokine Release from Bovine Alveolar Macrophages Based on RAGE-TLR4 Crosstalk[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(10): 2944-2952.

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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

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