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

作　　者：孙紫杭 鞠艺朵 徐若昕 陆诗雨 李建敏 殷姝宇 刘佃滨 布会敏 韦艳霞 SUN Zihang;JU Yiduo;XU Ruoxin;LU Shiyu;LI Jianmin;YIN Shuyu;LIU Tianbin;BU Huimin;WEI Yanxia(Jiangsu Provincial Key Laboratory of Immunity and Metabolism,Xuzhou Medical University,Xuzhou,Jiangsu 221004,China;School of Stomatology,Xuzhou Medical University;Department of Physiology,School of Basic Medical Sciences,Xuzhou Medical University)

机构地区：[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

出　　处：《徐州医科大学学报》2024年第11期796-799,共4页Journal of Xuzhou Medical University

基　　金：江苏省基础研究计划自然科学基金-面上项目(BK0190);徐州市重点研发计划(社会发展)项目-医药卫生面上(KC22122);江苏省大学生创新创业计划项目(202210313050Z);徐州市科技项目(KC21200)。

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。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 marrowderived 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 

分 类 号：R37[医药卫生—病原生物学]