title: Nrf2 and Snail-1 in the prevention of experimental liver fibrosis by caffeine.
abstract: OBJECTIVE
  To determine the molecular mechanisms involved in experimental hepatic fibrosis prevention by caffeine (CFA).
  
  
  METHODS
  Liver fibrosis was induced in Wistar rats by intraperitoneal thioacetamide or bile duct ligation and they were concomitantly treated with CFA (15 mg/kg per day). Fibrosis and inflammatory cell infiltrate were evaluated and classified by Knodell index. Inflammatory infiltrate was quantified by immunohistochemistry (anti-CD11b). Gene expression was analyzed by quantitative reverse transcription-polymerase chain reaction for collagen I (Col-1), connective tissue growth factor (CTGF), transforming growth factor β1 (TGF-β1), tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-1), IL-6, superoxide dismutase (SOD) and catalase (CAT). Activation of Nrf2 and Snail-1 was analyzed by Western-blot. TNF-α expression was proved by enzyme-linked immunosorbant assay, CAT activity was performed by zymography.
  
  
  RESULTS
  CFA treatment diminished fibrosis index in treated animals. The Knodell index showed both lower fibrosis and necroinflammation. Expression of profibrogenic genes CTGF, Col-1 and TGF-β1 and proinflammatory genes TNF-α, IL-6 and IL-1 was substantially diminished with CFA treatment with less CD11b positive areas. Significantly lower values of transcriptional factor Snail-1 were detected in CFA treated rats compared with cirrhotic rats without treatment; in contrast Nrf2 was increased in the presence of CFA. Expression of SOD and CAT was greater in animals treated with CFA showing a strong correlation between mRNA expression and enzyme activity.
  
  
  CONCLUSIONS
  Our results suggest that CFA inhibits the transcriptional factor Snail-1, down-regulating profibrogenic genes, and activates Nrf2 inducing antioxidant enzymes system, preventing inflammation and fibrosis.
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Daniela
authors.initials: D
authors.lastname: Gordillo-Bastidas
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Edén
authors.initials: E
authors.lastname: Oceguera-Contreras
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Adriana
authors.initials: A
authors.lastname: Salazar-Montes
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Jaime
authors.initials: J
authors.lastname: González-Cuevas
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Luis Daniel
authors.initials: LD
authors.lastname: Hernández-Ortega
authors.affiliation: Daniela Gordillo-Bastidas, Edén Oceguera-Contreras, Adriana Salazar-Montes, Jaime González-Cuevas, Luis Daniel Hernández-Ortega, Juan Armendáriz-Borunda, Institute of Molecular Biology in Medicine and Gene Therapy/CUCS, University of Guadalajara, Guadalajara, Jalisco 44100, México.
authors.forename: Juan
authors.initials: J
authors.lastname: Armendáriz-Borunda
mesh_terms: D000818:Animals; D001652:Bile Ducts / Q000601:surgery; D015415:Biomarkers / Q000378:metabolism; D039481:CD11b Antigen / Q000378:metabolism; D002110:Caffeine / Q000494:pharmacology*; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D024042:Collagen Type I / Q000235:genetics; D055513:Connective Tissue Growth Factor / Q000235:genetics; D019610:Cytoprotection; D005786:Gene Expression Regulation; D007375:Interleukin-1 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D008026:Ligation; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008106:Liver Cirrhosis, Experimental / Q000209:etiology / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000187:drug effects; D000071250:Snail Family Transcription Factors; D013482:Superoxide Dismutase / Q000235:genetics; D013853:Thioacetamide; D014157:Transcription Factors / Q000378:metabolism*; D053773:Transforming Growth Factor beta1 / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D015415:Biomarkers; C526316:CCN2 protein, rat; D039481:CD11b Antigen; D024042:Collagen Type I; D007375:Interleukin-1; D015850:Interleukin-6; D051267:NF-E2-Related Factor 2; C495637:Nfe2l2 protein, rat; D012333:RNA, Messenger; C000604450:Snai2 protein, rat; D000071250:Snail Family Transcription Factors; C508962:Tgfb1 protein, rat; D014157:Transcription Factors; D053773:Transforming Growth Factor beta1; D014409:Tumor Necrosis Factor-alpha; D013853:Thioacetamide; D055513:Connective Tissue Growth Factor; D002110:Caffeine; D002374:Catalase; D013482:Superoxide Dismutase
keywords: Antioxidant enzymes; Bile duct ligation; Caffeine; Liver fibrosis; Profibrogenic genes; Proinflammatory cytokines; Thioacetamide