肠道菌群代谢物DAT通过KLF4影响巨噬细胞IL-6产生
Gut microbial metabolite DAT affects the production of IL-6 in macrophages via KLF4
孙紫杭 1鞠艺朵 1徐若昕 1陆诗雨 1李建敏 1殷姝宇 1刘佃滨 2布会敏 3韦艳霞1
作者信息
- 1. 徐州医科大学江苏省免疫与代谢重点实验室,江苏 徐州 221004
- 2. 徐州医科大学口腔医学院
- 3. 徐州医科大学基础医学院生理教研室
- 折叠
摘要
目的 研究肠道菌群代谢产物脱氨基酪氨酸(DAT)对巨噬细胞白细胞介素-6(IL-6)的影响以及潜在的分子机制.方法 用脂多糖刺激骨髓来源巨噬细胞(BMDM),通过实时荧光定量PCR(q-PCR)和酶联免疫吸附试验(ELISA)检测DAT对BMDM IL-6、Krüppel样因子 4(KLF4)和P300/CBP相关因子(PCAF)mRNA和蛋白表达的影响.通过siRNA降低BMDM KLF4 表达,采用Western blot检测siRNA敲低KLF4 效率,通过q-PCR和ELISA检测IL-6、KLF4 mRNA和蛋白表达.结果 DAT可明显降低脂多糖刺激的BMDM上清液中IL-6 的表达(P<0.05),促进细胞KLF4 和PCAF基因转录;在敲低KLF4 的BMDM中,DAT降低IL-6 的作用被逆转.结论 在脂多糖刺激的BMDM中,DAT通过上调KLF4 减少IL-6 的生成.
Abstract
Objective To investigate the effect of the gut microbial metabolite deaminotyrosine(DAT)on the production of interleukin-6(IL-6)in macrophages and the underlying molecular mechanism.Methods Bone marrow-derived macrophages(BMDMs)were stimulated with lipopolysaccharide(LPS).The effect of DAT on the mRNA and protein expression of IL-6,Krüppel-like factor 4(KLF4),and P300/CBP-associated factor(PCAF)in BMDMs was detected by real-time quantitative PCR(q-PCR)and enzyme-linked immunosorbent assay(ELISA).The expression of KLF4 in BMDMs was knocked down by siRNA,and the efficiency of KLF4 knockdown by siRNA was detected by Western blot.The mRNA and protein expression of IL-6 and KLF4 were detected by q-PCR and ELISA.Results DAT significantly reduced IL-6 expression in the supernatant of LPS-stimulated BMDMs(P<0.05)and promoted the transcription of KLF4 and PCAF genes.In BMDMs with KLF4 knockdown,the inhibitory effect of DAT on IL-6 production was abolished.Conclusions In LPS-stimulated BMDMs,DAT reduces IL-6 production by upregulating KLF4 expression.
关键词
脱氨基酪氨酸/Krüppel样因子4/白细胞介素-6/骨髓来源巨噬细胞/肠道菌群/炎症反应Key words
deaminotyrosine/Krüppel-like factor 4/interleukin-6/bone marrow-derived macrophages/gut microbiota/inflammatory response引用本文复制引用
出版年
2024
NETL
NSTL
万方数据