title: MiR-9a-5p regulates proliferation and migration of hepatic stellate cells under pressure through inhibition of Sirt1.
abstract: OBJECTIVE
  To reveal the functions of microRNAs (miRNAs) with respect to hepatic stellate cells (HSCs) in response to portal hypertension.
  
  
  METHODS
  Primary rat HSCs were exposed to static water pressure (10 mmHg, 1 h) and the pressure-induced miRNA expression profile was detected by next-generation sequencing. Quantitative real-time polymerase chain reaction was used to verify the expression of miRNAs. A potential target of MiR-9a-5p was measured by a luciferase reporter assay and Western blot. CCK-8 assay and Transwell assay were used to detect the proliferation and migration of HSCs under pressure.
  
  
  RESULTS
  According to the profile, the expression of miR-9a-5p was further confirmed to be significantly increased after pressure overload in HSCs (3.70 ± 0.61 vs 0.97 ± 0.15, P = 0.0226), which resulted in the proliferation, migration and activation of HSCs. In vivo, the up-regulation of miR-9a-5p (2.09 ± 0.91 vs 4.27 ± 1.74, P = 0.0025) and the down-regulation of Sirt1 (2.41 ± 0.51 vs 1.13 ± 0.11, P = 0.0006) were observed in rat fibrotic liver with portal hypertension. Sirt1 was a potential target gene of miR-9a-5p. Through restoring the expression of Sirt1 in miR-9a-5p transfected HSCs on pressure overload, we found that overexpression of Sirt1 could partially abrogate the miR-9a-5p mediated suppression of the proliferation, migration and activation of HSCs.
  
  
  CONCLUSIONS
  Our results suggest that during liver fibrosis, portal hypertension may induce the proliferation, migration and activation of HSCs through the up-regulation of miR-9a-5p, which targets Sirt1.
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Qi
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Jiang-Feng
authors.initials: JF
authors.lastname: Hu
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Bao-Hai
authors.initials: BH
authors.lastname: Liu
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Chao-Qun
authors.initials: CQ
authors.lastname: Wu
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Hong-Yu
authors.initials: HY
authors.lastname: Yu
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Ding-Kang
authors.initials: DK
authors.lastname: Yao
authors.affiliation: Feng Qi, Bao-Hai Liu, Hong-Yu Yu, Ding-Kang Yao, Liang Zhu, Department of Gastroenterology, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zhu
mesh_terms: D000818:Animals; D001483:Base Sequence; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D020869:Gene Expression Profiling / Q000379:methods; D017930:Genes, Reporter; D055166:Hepatic Stellate Cells / Q000378:metabolism* / Q000473:pathology; D006975:Hypertension, Portal / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D008106:Liver Cirrhosis, Experimental / Q000150:complications; D008297:Male; D040542:Mechanotransduction, Cellular; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D017082:Portal Pressure; D061251:Primary Cell Culture; D051381:Rats; D017207:Rats, Sprague-Dawley; D060888:Real-Time Polymerase Chain Reaction; D056564:Sirtuin 1 / Q000235:genetics / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection
chemical_list: C582333:MIRN9 microRNA, rat; D035683:MicroRNAs; C488368:Sirt1 protein, rat; D056564:Sirtuin 1
keywords: Hepatic stellate cells; MicroRNA; Pressure; Sirt1; miR-9a-5p