title: Hes1, an important gene for activation of hepatic stellate cells, is regulated by Notch1 and TGF-β/BMP signaling.
abstract: OBJECTIVE
  To determine the role of Notch1 and Hes1 in regulating the activation of hepatic stellate cells (HSCs) and whether Hes1 is regulated by transforming growth factor (TGF)/bone morphogenetic protein (BMP) signaling.
  
  
  METHODS
  Immunofluorescence staining was used to detect the expression of desmin, glial fibrillary acidic protein and the myofibroblastic marker α-smooth muscle actin (α-SMA) after freshly isolated, normal rat HSCs had been activated in culture for different numbers of days (0, 1, 3, 7 and 10 d). The expression of α-SMA, collagen1α2 (COL1α2), Notch receptors (Notch1-4), and the Notch target genes Hes1 and Hey1 were analyzed by reverse transcriptase-polymerase chain reaction. Luciferase reporter assays and Western blot were used to study the regulation of α-SMA, COL1α1, COL1α2 and Hes1 by NICD1, Hes1, CA-ALK3, and CA-ALK5 in HSC-T6 cells. Moreover, the effects of inhibiting Hes1 function in HSC-T6 cells using a Hes1 decoy were also investigated.
  
  
  RESULTS
  The expression of Notch1 and Hes1 mRNAs was significantly down-regulated during the culture of freshly isolated HSCs. In HSC-T6 cells, Notch1 inhibited the promoter activities of α-SMA, COL1α1 and COL1α2. On the other hand, Hes1 enhanced the promoter activities of α-SMA and COL1α2, and this effect could be blocked by inhibiting Hes1 function with a Hes1 decoy. Furthermore, co-transfection of pcDNA3-CA-ALK3 (BMP signaling activin receptor-like kinase 3) and pcDNA3.1-NICD1 further increased the expression of Hes1 compared with transfection of either vector alone in HSC-T6 cells, while pcDNA3-CA-ALK5 (TGF-β signaling activin receptor-like kinase 5) reduced the effect of NICD1 on Hes1 expression.
  
  
  CONCLUSIONS
  Selective interruption of Hes1 or maintenance of Hes1 at a reasonable level decreases the promoter activities of α-SMA and COL1α2, and these conditions may provide an anti-fibrotic strategy against hepatic fibrosis.
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhang
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Yan-Qiong
authors.initials: YQ
authors.lastname: Zhang
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Wen-Bing
authors.initials: WB
authors.lastname: Ai
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Qing-Ting
authors.initials: QT
authors.lastname: Hu
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Qiao-Juan
authors.initials: QJ
authors.lastname: Zhang
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Lin-Yan
authors.initials: LY
authors.lastname: Wan
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Xiao-Lian
authors.initials: XL
authors.lastname: Wang
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Chang-Bai
authors.initials: CB
authors.lastname: Liu
authors.affiliation: Kai Zhang, Yan-Qiong Zhang, Qing-Ting Hu, Qiao-Juan Zhang, Lin-Yan Wan, Xiao-Lian Wang, Chang-Bai Liu, Jiang-Feng Wu, Medical College, China Three Gorges University, Yichang 443002, Hubei Province, China.
authors.forename: Jiang-Feng
authors.initials: JF
authors.lastname: Wu
mesh_terms: D000199:Actins / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D052005:Bone Morphogenetic Protein Receptors, Type I / Q000235:genetics / Q000378:metabolism; D019485:Bone Morphogenetic Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D055166:Hepatic Stellate Cells / Q000378:metabolism* / Q000473:pathology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D008103:Liver Cirrhosis / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D058628:Myofibroblasts / Q000378:metabolism / Q000473:pathology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D051881:Receptor, Notch1 / Q000235:genetics / Q000378:metabolism*; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013997:Time Factors; D000072056:Transcription Factor HES-1; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000378:metabolism*
chemical_list: D000199:Actins; D051792:Basic Helix-Loop-Helix Proteins; D015415:Biomarkers; D052005:Bone Morphogenetic Protein Receptors, Type I; D019485:Bone Morphogenetic Proteins; D024042:Collagen Type I; D018398:Homeodomain Proteins; D017346:Protein Serine-Threonine Kinases; D012333:RNA, Messenger; D051881:Receptor, Notch1; D000077293:Receptor, Transforming Growth Factor-beta Type I; D018125:Receptors, Transforming Growth Factor beta; D000072056:Transcription Factor HES-1; D016212:Transforming Growth Factor beta; C489735:Hes1 protein, rat; C497763:Notch1 protein, rat; C498862:Acta2 protein, rat; C000625843:Tgfbr1 protein, rat
keywords: Hepatic fibrosis; Hepatic stellate cells; Hes1; Notch1; TGF-β/BMP