Lingjian Kong1, Jing Chen2, Xiaoli Ji3, Qian Qin4, Huiyu Yang5, Dan Liu5, Deliang Li5, Meiling Sun6. 1. Department of Gastroenterology, The First Affiliated Hospital of Zhengzhou University, No.1 Jianshe East Road, Henan Province, Zhengzhou, 450052, PR China. konggewww@163.com. 2. Department of Gastroenterology, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang, 150086, PR China. 3. Department of Intervention, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, 450052, PR China. 4. Physical Examination Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, 450052, PR China. 5. Department of Gastroenterology, The First Affiliated Hospital of Zhengzhou University, No.1 Jianshe East Road, Henan Province, Zhengzhou, 450052, PR China. 6. Department of Gastroenterology, ZhuJiang Hospital of Southern Medical University, Guangzhou, Guangdong Province, 510280, PR China.
Abstract
BACKGROUND: Alcohol-induced intestinal dysbiosis disrupts and inflammatory responses are essential in the development of alcoholic fatty liver disease (AFLD). Here, we investigated the effects of Fmo5 on changes in enteric microbiome composition in a model of AFLD and dissected the pathogenic role of Fmo5 in AFLD-induced liver pathology. METHODS: The expression profile data of GSE8006 and GSE40334 datasets were downloaded from the GEO database. The WGCNA approach allowed us to investigate the AFLD-correlated module. DEGs were used to perform KEGG pathway enrichment analyses. Four PPI networks were constructed using the STRING database and visualized using Cytoscape software. The Cytohubba plug-in was used to identify the hub genes. Western blot and immunohistochemistry assays were used to detect protein expression. ELISA assay was used to detect the levels of serum inflammatory cytokines. Lipid droplets in the cytoplasm were observed using Oil Red O staining. Apoptosis was detected using a TUNEL assay and flow cytometry analysis. ROS levels were detected using flow cytometry analysis. Nuclear translocation of NF-κB p65 was observed using immunofluorescence staining. Co-immunoprecipitation was used to detect the co-expression of PPARα and Fmo5 in L02 cells. 16S rDNA sequencing defined the bacterial communities in mice with AFLD. RESULTS: Fmo5 is a key DEG and is closely associated with the gut microbiota and PPAR signaling pathway. Gut microbiome function in AFLD was significantly related to the PPAR signaling pathway. AFLD induced shifts in various bacterial phyla in the cecum, including a reduction in Bacteroidetes and increased Firmicutes. Fmo5 and PPARα co-expression in cell and animal models with AFLD, which decreased significantly. Silencing of Fmo5 and PPARα aggravated the functions of AFLD inducing apoptosis and inflammatory response, promoting liver injury, and activating the NF-κB signaling pathway in vivo and in vitro. The NF-κB inhibitor abolished the functions of silencing of Fmo5 and PPARα promoting AFLD-induced apoptosis, inflammatory response, and liver injury. CONCLUSION: Our data indicated that the co-expression of Fmo5 and PPARα was involved in AFLD-related gut microbiota composition and alleviated AFLD-induced liver injury, apoptosis, and inflammatory response by inhibiting the nuclear translocation of NF-κB p65 to inhibit the NF-κB signaling pathway.
BACKGROUND:Alcohol-induced intestinal dysbiosis disrupts and inflammatory responses are essential in the development of alcoholic fatty liver disease (AFLD). Here, we investigated the effects of Fmo5 on changes in enteric microbiome composition in a model of AFLD and dissected the pathogenic role of Fmo5 in AFLD-induced liver pathology. METHODS: The expression profile data of GSE8006 and GSE40334 datasets were downloaded from the GEO database. The WGCNA approach allowed us to investigate the AFLD-correlated module. DEGs were used to perform KEGG pathway enrichment analyses. Four PPI networks were constructed using the STRING database and visualized using Cytoscape software. The Cytohubba plug-in was used to identify the hub genes. Western blot and immunohistochemistry assays were used to detect protein expression. ELISA assay was used to detect the levels of serum inflammatory cytokines. Lipid droplets in the cytoplasm were observed using Oil Red O staining. Apoptosis was detected using a TUNEL assay and flow cytometry analysis. ROS levels were detected using flow cytometry analysis. Nuclear translocation of NF-κB p65 was observed using immunofluorescence staining. Co-immunoprecipitation was used to detect the co-expression of PPARα and Fmo5 in L02 cells. 16S rDNA sequencing defined the bacterial communities in mice with AFLD. RESULTS:Fmo5 is a key DEG and is closely associated with the gut microbiota and PPAR signaling pathway. Gut microbiome function in AFLD was significantly related to the PPAR signaling pathway. AFLD induced shifts in various bacterial phyla in the cecum, including a reduction in Bacteroidetes and increased Firmicutes. Fmo5 and PPARα co-expression in cell and animal models with AFLD, which decreased significantly. Silencing of Fmo5 and PPARα aggravated the functions of AFLD inducing apoptosis and inflammatory response, promoting liver injury, and activating the NF-κB signaling pathway in vivo and in vitro. The NF-κB inhibitor abolished the functions of silencing of Fmo5 and PPARα promoting AFLD-induced apoptosis, inflammatory response, and liver injury. CONCLUSION: Our data indicated that the co-expression of Fmo5 and PPARα was involved in AFLD-related gut microbiota composition and alleviated AFLD-induced liver injury, apoptosis, and inflammatory response by inhibiting the nuclear translocation of NF-κB p65 to inhibit the NF-κB signaling pathway.
Authors: F J Oliver; J Ménissier-de Murcia; C Nacci; P Decker; R Andriantsitohaina; S Muller; G de la Rubia; J C Stoclet; G de Murcia Journal: EMBO J Date: 1999-08-16 Impact factor: 11.598
Authors: Flora Scott; Sandra G Gonzalez Malagon; Brett A O'Brien; Diede Fennema; Sunil Veeravalli; Clarissa R Coveney; Ian R Phillips; Elizabeth A Shephard Journal: Drug Metab Dispos Date: 2017-06-23 Impact factor: 3.922
Authors: Yi Duan; Cristina Llorente; Sonja Lang; Katharina Brandl; Huikuan Chu; Lu Jiang; Richard C White; Thomas H Clarke; Kevin Nguyen; Manolito Torralba; Yan Shao; Jinyuan Liu; Adriana Hernandez-Morales; Lauren Lessor; Imran R Rahman; Yukiko Miyamoto; Melissa Ly; Bei Gao; Weizhong Sun; Roman Kiesel; Felix Hutmacher; Suhan Lee; Meritxell Ventura-Cots; Francisco Bosques-Padilla; Elizabeth C Verna; Juan G Abraldes; Robert S Brown; Victor Vargas; Jose Altamirano; Juan Caballería; Debbie L Shawcross; Samuel B Ho; Alexandre Louvet; Michael R Lucey; Philippe Mathurin; Guadalupe Garcia-Tsao; Ramon Bataller; Xin M Tu; Lars Eckmann; Wilfred A van der Donk; Ry Young; Trevor D Lawley; Peter Stärkel; David Pride; Derrick E Fouts; Bernd Schnabl Journal: Nature Date: 2019-11-13 Impact factor: 49.962