摘要
目的研究脱氧胆酸(DCA)对肠上皮细胞趋化因子MCP-1表达的诱导作用及机制。方法以不同浓度DCA刺激小鼠肠上皮细胞,荧光定量PCR和ELISA法分别检测细胞MCP-1 mRNA、蛋白表达水平;利用siRNA技术或受体拮抗剂抑制相关胆汁酸受体,荧光定量PCR检测DCA对诱导MCP-1表达的影响;进一步分别阻断NF-κB和丝裂原活化蛋白激酶(MAPK)信号传导通路,检测其对DCA诱导MCP-1表达的影响,并通过蛋白印迹验证DCA对肠上皮细胞NF-κB、MAPK相关信号通路的活化作用。结果DCA可剂量依赖性地诱导小鼠肠上皮细胞表达MCP-1,该作用主要由胆汁酸受体M2-mAchR介导;特异性的NF-κB、JNK或ERK抑制剂可明显抑制DCA诱导的MCP-1表达(P<0.001),同时DCA处理肠上皮细胞可显著上调ERK1/2、JNK和IκB的磷酸化水平。结论高水平DCA可诱导肠上皮细胞产生MCP-1,这可能是其引起肠道巨噬细胞浸润、促进肠道炎症反应的重要作用机制之一。
Objective To investigate the mechanism by which the chemokine monocyte chemotactic protein 1(MCP-1)expression is induced by deoxycholic acid(DCA)in murine intestinal epithelial cells.Methods Murine intestinal epi-thelial cells were stimulated with different concentrations of DCA,and MCP-1 mRNA and protein expression levels were detected by real-time PCR and ELISA respectively.Bile acid receptors were inhibited by either siRNA or related receptor antagonist while DCA-induced MCP-1 expression was evaluated by real-time PCR.Furthermore,NF-κB and mitogen-acti-vated protein kinases(MAPKs)signaling pathways were respectively blocked to explore the corresponding effects on the MCP-1 expression induced by DCA.Western blotting was applied to confirm the activation of NF-κB and MAPKs-related signaling pathways.Results DCA could dose-dependently induce the expression of MCP-1 in murine intestinal epithelial cells and this effect was mainly mediated by bile acid receptor-M2 muscarinic acetylcholine receptor(M2-mAchR).Specif-ic inhibitors of nuclear factor(NF)-κB,c-Jun N-terminal kinase(JNK)or extracellular regulated kinase(ERK)significant-ly decreased MCP-1 expression induced by DCA(P<0.001).Meanwhile,DCA could dramatically upregulate the phos-phorylation levels of ERK1/2,JNK and IκB.Conclusion High levels of DCA can effectively induce MCP-1 expression in intestinal epithelial cells,which may be one of the most important mechanisms that cause intestinal macrophage infiltration and promote intestinal inflammation.
作者
王岭玉
田春艳
吴瑾
WANG Ling yu;TIAN Chun yan;WU Jin(Xinhua Hospital,School of Medicine,Shanghai Jiaotong University,Shanghai 200092,China;Shanghai Institute for Pediatric Research,Shanghai 200092,China;Shanghai Key Laboratory of Pediatric Gastroenterology and Nutrition,Shanghai 200092,China;Institute of Lifeomics,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 102206,China)
出处
《军事医学》
CAS
北大核心
2019年第12期897-901,共5页
Military Medical Sciences
基金
上海市自然科学基金(16ZR1428500)
上海市卫生健康委员会资助项目(201940219).
关键词
炎症性肠病
脱氧胆酸
高脂饮食
单核细胞趋化蛋白1
肠上皮细胞
inflammatory bowel disease
deoxycholic acid
high fat diet
monocyte chemotactic protein-1
intestinal epithelial cells
作者简介
王岭玉,女,硕士研究生,研究方向:炎症性肠病的发病机制;通讯作者:吴瑾,Tel:02125076443,Email:wujin@xinhuamed.com.cn
