title: Hydrodynamic-based in vivo transfection of retinoic X receptor-alpha gene can enhance vitamin A-induced attenuation of liver fibrosis in mice.
abstract: OBJECTIVE
  In hepatic stellate cells isolated from rat fibrotic livers, the amount of retinoid X receptor-alpha (RXR-alpha) mRNA is greatly reduced. However, the effectiveness of retinoids in the treatment of liver fibrosis is controversial. We hypothesized that increasing the expression levels of RXR-alpha in livers will improve the response of liver fibrosis to retinoids treatment.
  
  
  METHODS
  pTracer-CMV2 vector harboring both green fluorescent protein and RXR-alpha genes was given to mice with carbon tetrachloride (CCl(4))-induced liver fibrosis, by hydrodynamic-based in vivo transfection. Vitamin A was simultaneously administered to the mice. Sirius red staining and measurement of hydroxyproline content were performed to evaluate liver fibrosis. The incorporation of 5-bromo-2-deoxyribouridine (BrdU) was carried out to determine liver cell proliferation.
  
  
  RESULTS
  Successful transfection and expression of exogenous RXR-alpha gene in the liver was determined by observance of green fluorescence under a confocal microscope, and detection of RXR-alpha protein by immunohistochemistry. Hepatic fibrosis, evaluated by both Sirius red staining with image analysis and quantity of hydroxyproline in livers of RXR-alpha-transfected group, tapered off remarkably. The hydroxyproline content and Sirius red-positive staining area on liver sections from RXR-alpha-transfected mice decreased by 34.3% and 54.63%, respectively, compared with the control group receiving empty vector. The labeling index of BrdU in non-parenchymal cells was much lower in livers from the RXR-alpha-transfected group than that of empty vector-transfected group.
  
  
  CONCLUSIONS
  Hydrodynamic-based in vivo transfection of the RXR-alpha gene can enhance the vitamin A-induced attenuation of liver fibrosis in mice. One of the possible mechanisms of action for this gene treatment is inhibition of non-parenchymal cell proliferation mainly composed of hepatic stellate cells in fibrotic livers.
authors.affiliation: Department of Pathology, Shanghai Medical College, Fudan University, Shanghai, China.
authors.forename: Chen
authors.initials: C
authors.lastname: Chen
authors.forename: Jinsheng
authors.initials: J
authors.lastname: Zhang
authors.forename: Jinjun
authors.initials: J
authors.lastname: Li
authors.forename: Jin
authors.initials: J
authors.lastname: Huang
authors.forename: Chen
authors.initials: C
authors.lastname: Yang
authors.forename: Guangcun
authors.initials: G
authors.lastname: Huang
authors.forename: Jianying
authors.initials: J
authors.lastname: Shi
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D022781:Hepatocytes / Q000378:metabolism; D007150:Immunohistochemistry; D008106:Liver Cirrhosis, Experimental / Q000473:pathology* / Q000628:therapy*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D018613:Microscopy, Confocal; D011336:Probability; D012333:RNA, Messenger / Q000378:metabolism; D047488:Retinoid X Receptors / Q000235:genetics*; D012680:Sensitivity and Specificity; D014162:Transfection; D014801:Vitamin A / Q000494:pharmacology*
chemical_list: D012333:RNA, Messenger; D047488:Retinoid X Receptors; D014801:Vitamin A