title: Trichostatin A inhibits transforming growth factor-β-induced reactive oxygen species accumulation and myofibroblast differentiation via enhanced NF-E2-related factor 2-antioxidant response element signaling.
abstract: Trichostatin A (TSA) has been shown to prevent fibrosis in vitro and in vivo. The present study aimed at investigating the role of reactive oxygen species (ROS) scavenging by TSA on transforming growth factor-β (TGF-β)-induced myofibroblast differentiation of corneal fibroblasts in vitro. Human immortalized corneal fibroblasts were treated with TGF-β in the presence of TSA, the NAD(P)H oxidase inhibitor diphenyleneiodonium (DPI), the antioxidant N-acetyl-cysteine (NAC), the NF-E2-related factor 2-antioxidant response element (Nrf2-ARE) activator sulforaphane, or small interfering RNA. Myofibroblast differentiation was assessed by α-smooth muscle actin (α-SMA) expression, F-actin bundle formation, and collagen gel contraction. ROS, H(2)O(2), intracellular glutathione (GSH) level, cellular total antioxidant capacity, and the activation of Nrf2-ARE signaling were determined with various assays. Treatment with TSA and the Nrf2-ARE activator resulted in increased inhibition of the TGF-β-induced myofibroblast differentiation as compared with treatment with DPI or NAC. Furthermore, TSA also decreased cellular ROS and H(2)O(2) accumulation induced by TGF-β, whereas it elevated intracellular GSH level and cellular total antioxidant capacity. In addition, TSA induced Nrf2 nuclear translocation and up-regulated the expression of Nrf2-ARE downstream antioxidant genes, whereas Nrf2 knockdown by RNA interference blocked the inhibition of TSA on myofibroblast differentiation. In conclusion, this study provides the first evidence implicating that TSA inhibits TGF-β-induced ROS accumulation and myofibroblast differentiation via enhanced Nrf2-ARE signaling.
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of Medical Sciences, Qingdao, China.
authors.forename: Lingling
authors.initials: L
authors.lastname: Yang
authors.forename: Mingli
authors.initials: M
authors.lastname: Qu
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.forename: Haoyun
authors.initials: H
authors.lastname: Duan
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
authors.forename: Ye
authors.initials: Y
authors.lastname: Wang
authors.forename: Weiyun
authors.initials: W
authors.lastname: Shi
authors.forename: Patrik
authors.initials: P
authors.lastname: Danielson
authors.forename: Qingjun
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000199:Actins / Q000378:metabolism; D061745:Antioxidant Response Elements / Q000187:drug effects; D000975:Antioxidants / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D003094:Collagen / Q000378:metabolism; D003315:Cornea / Q000187:drug effects / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000037:antagonists & inhibitors* / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology*; D017879:Isothiocyanates; D058628:Myofibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors; D051267:NF-E2-Related Factor 2 / Q000378:metabolism*; D009861:Onium Compounds / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013454:Sulfoxides; D013861:Thiocyanates / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: C541116:ACTA2 protein, human; D000199:Actins; D000975:Antioxidants; D006877:Hydroxamic Acids; D017879:Isothiocyanates; D051267:NF-E2-Related Factor 2; D009861:Onium Compounds; D017382:Reactive Oxygen Species; D013454:Sulfoxides; D013861:Thiocyanates; D016212:Transforming Growth Factor beta; C012589:trichostatin A; C007517:diphenyleneiodonium; D003094:Collagen; D006861:Hydrogen Peroxide; D019255:NADPH Oxidases; C016766:sulforaphane; D005978:Glutathione