title: Givinostat inhibition of hepatic stellate cell proliferation and protein acetylation.
abstract: OBJECTIVE
  To explore the effect of the histone deacetylase inhibitor givinostat on proteins related to regulation of hepatic stellate cell proliferation.
  
  
  METHODS
  The cell counting kit-8 assay and flow cytometry were used to observe changes in proliferation, apoptosis, and cell cycle in hepatic stellate cells treated with givinostat. Western blot was used to observe expression changes in p21, p57, CDK4, CDK6, cyclinD1, caspase-3, and caspase-9 in hepatic stellate cells exposed to givinostat. The scratch assay was used to analyze the effect of givinostat on cell migration. Effects of givinostat on the reactive oxygen species profile, mitochondrial membrane potential, and mitochondrial permeability transition pore opening in JS-1 cells were observed by laser confocal microscopy.
  
  
  RESULTS
  Givinostat significantly inhibited JS-1 cell proliferation and promoted cell apoptosis, leading to cell cycle arrest in G0/G1 phases. Treatment with givinostat downregulated protein expression of CDK4, CDK6, and cyclin D1, whereas expression of p21 and p57 was significantly increased. The givinostat-induced apoptosis of hepatic stellate cells was mainly mediated through p38 and extracellular signal-regulated kinase 1/2. Givinostat treatment increased intracellular reactive oxygen species production, decreased mitochondrial membrane potential, and promoted mitochondrial permeability transition pore opening. Acetylation of superoxide dismutase (acetyl K68) and nuclear factor-κB p65 (acetyl K310) was upregulated, while there was no change in protein expression. Moreover, the notable beneficial effect of givinostat on liver fibrosis was also confirmed in the mouse models.
  
  
  CONCLUSIONS
  Givinostat has antifibrotic activities via regulating the acetylation of nuclear factor-κB and superoxide dismutase 2, thus inhibiting hepatic stellate cell proliferation and inducing apoptosis.
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Yu-Gang
authors.initials: YG
authors.lastname: Wang
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Xu
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Ting
authors.initials: T
authors.lastname: Wang
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Jue
authors.initials: J
authors.lastname: Wei
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Wen-Ying
authors.initials: WY
authors.lastname: Meng
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Na
authors.initials: N
authors.lastname: Wang
authors.affiliation: Yu-Gang Wang, Ling Xu, Ting Wang, Jue Wei, Wen-Ying Meng, Na Wang, Min Shi, Department of Gastroenterology, Shanghai Tongren Hospital, Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200336, China.
authors.forename: Min
authors.initials: M
authors.lastname: Shi
mesh_terms: D000107:Acetylation; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002219:Carbamates / Q000494:pharmacology*; D002251:Carbon Tetrachloride; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects*; D056486:Chemical and Drug Induced Liver Injury / Q000209:etiology / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004305:Dose-Response Relationship, Drug; D055166:Hepatic Stellate Cells / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D056572:Histone Deacetylase Inhibitors / Q000494:pharmacology*; D008106:Liver Cirrhosis, Experimental / Q000139:chemically induced / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D008297:Male; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D015398:Signal Transduction / Q000187:drug effects; D013482:Superoxide Dismutase / Q000378:metabolism; D013997:Time Factors; D000099156:Superoxide Dismutase 2
chemical_list: D051017:Apoptosis Regulatory Proteins; D002219:Carbamates; D002251:Carbon Tetrachloride; D018797:Cell Cycle Proteins; D056572:Histone Deacetylase Inhibitors; D016328:NF-kappa B; D013482:Superoxide Dismutase; D000099156:Superoxide Dismutase 2; C575255:givinostat
keywords: Givinostat; Hepatic stellate cells; Histone deacetylase inhibitor; Nuclear factor-κB; Superoxide dismutase