临床肝胆病杂志2025，Vol.41Issue(12)：2536-2544,9.DOI:10.12449/JCH251215
扶正化瘀胶囊对代谢相关脂肪性肝病肝纤维化小鼠模型疤痕相关巨噬细胞的调控作用及机制分析
Regulatory role and mechanism of Fuzheng Huayu Capsule on scar-associated macrophages in a mouse model of liver fibrosis in metabolic associated fatty liver disease
摘要
Abstract
Objective To investigate the molecular mechanism of Fuzheng Huayu Capsule in improving liver fibrosis in metabolic associated fatty liver disease by regulating scar-associated macrophages(SAMs).Methods A total of 24 C57 mice were randomly divided into control group(Con group),Model group,and Fuzheng Huayu Capsule group(FZHY group).Mice were given a high-fat diet and intraperitoneal injection of CCl4 for six weeks to establish a model of liver fibrosis in metabolic associated fatty liver disease.The drug was given by gavage for 5 consecutive weeks since week 2 of modeling.FZHY group was administered Fuzheng Huayu Capsule via oral gavage,while the Con and Model groups received an equal volume of saline solution via oral gavage.The serum levels of liver enzymes were measured,as well as the levels of triglyceride(TG)and hydroxyproline(Hyp)in the liver.HE staining and picrosirius red staining were used to observe liver tissue.Three liver tissue samples were collected from the Model group and the FZHY group,and transcriptome sequencing was performed to investigate the molecular mechanism of Fuzheng Huayu Capsule in improving liver fibrosis in metabolic associated fatty liver disease.Western blot and RT-qPCR were used to measure the protein and/or mRNA expression levels of SAM markers(CD9 and triggering receptor expressed on myeloid cells 2[TREM2]),profibrogenic genes(transforming growth factor-β1[TGFβ1],platelet-derived growth factor subunit beta[PDGFβ],and TNF superfamily member 12[TNFSF12]),and the upstream regulator activating transcription factor 3(ATF3)in liver tissue.The serum containing Fuzheng Huayu Capsule was used for the intervention of pro-inflammatory bone marrow-derived macrophages(BMDMs)induced by lipopolysaccharide and TGF-β1,and immunofluorescence assay,Western blot,and RT-qPCR were used to measure the expression levels of TREM2 and ATF3.The independent-samples t test was used for comparison of continuous data between two groups,and a one-way analysis of variance was used for comparison between multiple groups,while the least significant difference t-test was used for further comparison between two groups.Results Compared with the Model group,the FZHY group had significant reductions in the levels of liver enzymes,the levels of TG and Hyp in the liver,NAS score,and Sirius Red staining-positive area(all P<0.01).The RNA-seq analysis showed that differentially expressed genes were mainly enriched in chemokine signaling pathways,and Fuzheng Huayu Capsule significantly downregulated the expression of CCL2,CX3CL1,and CX3CR1(all P<0.01).Fuzheng Huayu Capsule significantly reduced the protein and mRNA expression levels of CD9,TREM2,and ATF3 in liver tissue(all P<0.05).In vitro,Fuzheng Huayu Capsule significantly reduced the mRNA expression levels of TGFβ1,PDGFβ,and TNFSF12 in liver tissue(all P<0.01).Fuzheng Huayu Capsule also attenuated TREM2 fluorescence intensity in pro-inflammatory BMDMs and significantly reduced the mRNA and protein expression levels of ATF3(all P<0.05).Conclusion Fuzheng Huayu Capsule has a marked therapeutic effect on mice with liver fibrosis in metabolic associated fatty liver disease,possibly by downregulating the expression of ATF3 and inhibiting SAMs.关键词
代谢相关脂肪性肝病/肝纤维化/扶正化瘀胶囊/巨噬细胞/转录激活因子3/小鼠,近交C57BLKey words
Metabolic Associated Fatty Liver Disease/Liver Fibrosis/Fuzheng Huayu Capsules/Macrophages/Activating Transcription Factor 3/Mice,Inbred C57BL引用本文复制引用
邹晨,林儒焘,徐笑,黄倩,刘平,胡义扬,辛鑫,冯琴..扶正化瘀胶囊对代谢相关脂肪性肝病肝纤维化小鼠模型疤痕相关巨噬细胞的调控作用及机制分析[J].临床肝胆病杂志,2025,41(12):2536-2544,9.基金项目
上海市青年科技英才扬帆计划项目(23YF1448200) Shanghai Youth Technology Talents Sailing Program(23YF1448200) （23YF1448200）
