中国中医药信息杂志2025，Vol.32Issue(6)：128-134,7.DOI:10.19879/j.cnki.1005-5304.202412662
基于肥大细胞外泌体miR-34c对肠屏障功能的影响探讨清毒汤治疗慢加急性肝衰竭分子机制
Exploration on the Molecular Mechanism of Qingdu Decoction in the Treatment of Acute-on-Chronic Liver Failure Based on the Effects of miR-34c in Mast Cell Exosomes on Intestinal Barrier Function
摘要
Abstract
Objective To observe the effects of Qingdu Decoction on intestinal barrier function by regulating the miR-34c-mediated signaling pathway of human mast cells-derived exosomes;To explore its molecular mechanism in the treatment of acute-on-chronic liver failure(ACLF).Methods An inflammatory model was established by treating human mast cells with 1 μg/mL lipopolysaccharide.The mast cells were over-expressed with miR-34c using lentiviral transfection,and were divided into normal group,model group and miR-34c over-expression group.Exosomes were extracted from each group of mast cells.Human intestinal epithelial cells were divided into normal group,normal exosome group,model exosome group,miR-34c over-expression exosome group and Qingdu Decoction group(miR-34c overexpression exosomes+Qingdu Decoction),and corresponding treatment was given.The dual luciferase gene report confirmed the targeting relationship between miR-34c and myeloma associated oncogene zinc finger protein(MAZ).RT-qPCR was used to detect the expression of miR-34c in mast cells and intestinal epithelial cells,and Western blot was used to detect the expressions of MAZ,Occludin and ZO-1 protein.Results Compared with the normal group,the expression of miR-34c in mast cells treated with lipopolysaccharide and transfected with lentivirus significantly increased(P<0.01).The results of the dual luciferase reporter gene experiment showed that miR-34c could target and inhibit MAZ expression.The RT-qPCR results showed that compared with the normal group,the miR-34c expression in intestinal epithelial cells significantly increased in the model exosome group and miR-34c over-expression exosome group(P<0.05,P<0.01),while the miR-34c expression in Qingdu Decoction group significantly decreased compared with the miR-34c over-expression exosome group(P<0.01).The Western blot results showed that compared with the normal group,the expressions of MAZ,Occludin and ZO-1 proteins in the model exosome group and miR-34c over-expressing exosome group were significantly reduced(P<0.05,P<0.01),while the expressions of MAZ,Occludin and ZO-1 proteins in Qingdu Decoction group significantly increased compared with the miR-34c over-expressing exosome group(P<0.05).Conclusion The treatment of ACLF with Qingdu Decoction may be related to the regulation of the miR-34c-mediated signaling pathway in mast cell-derived exosomes and the up-regulation of intestinal epithelial tight junction protein expression.关键词
慢加急性肝衰竭/清毒汤/外泌体/miR-34c/肠上皮细胞/紧密连接蛋白Key words
acute-on-chronic liver failure/Qingdu Decoction/exosomes/miR-34c/intestinal epithelial cells/tight junction protein分类
医药卫生引用本文复制引用
杨佳璇,陈月,师娟,高连印..基于肥大细胞外泌体miR-34c对肠屏障功能的影响探讨清毒汤治疗慢加急性肝衰竭分子机制[J].中国中医药信息杂志,2025,32(6):128-134,7.基金项目
国家自然科学基金面上项目(82474301) （82474301）
北京市自然科学基金面上项目(7222266) （7222266）
