title: Lazaroid U-74389G inhibits the osteoblastic differentiation of IL-1β-indcued aortic valve interstitial cells through glucocorticoid receptor and inhibition of NF-κB pathway.
abstract: BACKGROUND
  Aortic valve calcification is characterized as the active process of aortic valve interstitial cells (AVICs), and considered as an inflammatory disease. As an antioxidant, the anti-inflammatory activity of Lazaroid has been exhibited in various models. We hypothesized that Lazaroid U-74389G would inhibit the osteoblastic differentiation of AVICs induced by IL-1β.
  
  
  METHODS
  Normal tricuspid aortic valve leaflets were collected from patients with acute aortic dissection (Type A) undergoing the Bentall procedure. AVICs were isolated and stimulated with IL-1β in presence or absence of U-74389G in culture. Cell lysates were analyzed for osteogenic markers and nuclear factor-κB using real-time PCR and Immunoblotting. Culture media was analyzed for IL-6 and IL-8 with enzyme-linked immunosorbent assay. Alizarin Red Staining was adopted to demonstrate the calcium deposition.
  
  
  RESULTS
  The expression of alkaline phosphatase and bone morphogenetic protein, accompanied by the production of IL-6 and IL-8, was up-regulated in response to IL-1β and was inhibited by the addition of U-74389G. The NF-κB pathway was activated by IL-1β and involved in the suppression of U-74389G on osteoblastic differentiation in AVICs. The negative effects of U-74389G on ostengenic gene expression and mineralization of AVICs were blocked by glucocorticoid receptor antagonist mifepristone and the NF-κB inhibitor Bay 11-7082.
  
  
  CONCLUSIONS
  U-74389G inhibits the pro-osteogenic response to IL-1β stimulation in AVICs. The osteoblastic differentiation and mineralization of AVICs were inhabited by U-74389G though the modulation of NF-κB activation, and this pathway could be potential therapeutic targets for medical treatment of calcified aortic valve disease.
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China; Department of Cardiovascular surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China. Electronic address: sfq1010@126.com.
authors.forename: Fuqiang
authors.initials: F
authors.lastname: Sun
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: shijiawei@21cn.com.
authors.forename: Jiawei
authors.initials: J
authors.lastname: Shi
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: 3975840@qq.com.
authors.forename: Si
authors.initials: S
authors.lastname: Chen
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: rivaldodc@163.com.
authors.forename: Cheng
authors.initials: C
authors.lastname: Deng
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: 40037949@qq.com.
authors.forename: Xingjian
authors.initials: X
authors.lastname: Hu
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: lihuadong.416@163.com.
authors.forename: Huadong
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: ligeng66@126.com.
authors.forename: Geng
authors.initials: G
authors.lastname: Li
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: liuyi2268bo@sina.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Cardiovascular surgery, Union Hospital, Huazhong University of Science and Technology, Wuhan 430022, China. Electronic address: dongnianguo@hotmail.com.
authors.forename: Nianguo
authors.initials: N
authors.lastname: Dong
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis; D001021:Aortic Valve / Q000166:cytology / Q000473:pathology; D001024:Aortic Valve Stenosis / Q000473:pathology; D019485:Bone Morphogenetic Proteins / Q000096:biosynthesis; D002114:Calcinosis / Q000473:pathology; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D053583:Interleukin-1beta / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism; D016209:Interleukin-8 / Q000378:metabolism; D008297:Male; D015735:Mifepristone / Q000494:pharmacology; D032389:Myocytes, Smooth Muscle / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D010006:Osteoblasts / Q000166:cytology; D010012:Osteogenesis / Q000187:drug effects*; D010766:Phosphorylation / Q000187:drug effects; D011281:Pregnatrienes / Q000494:pharmacology*; D011965:Receptors, Glucocorticoid / Q000037:antagonists & inhibitors / Q000378:metabolism*; D013450:Sulfones / Q000494:pharmacology; D051996:Transcription Factor RelA / Q000037:antagonists & inhibitors*; D014261:Tricuspid Valve / Q000166:cytology
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; D019485:Bone Morphogenetic Proteins; C508606:CXCL8 protein, human; C508585:IL1B protein, human; C508600:IL6 protein, human; D018836:Inflammation Mediators; D053583:Interleukin-1beta; D015850:Interleukin-6; D016209:Interleukin-8; D009570:Nitriles; D011281:Pregnatrienes; C498143:RELA protein, human; D011965:Receptors, Glucocorticoid; D013450:Sulfones; D051996:Transcription Factor RelA; C074965:U 74389F; D015735:Mifepristone; D000469:Alkaline Phosphatase
keywords: Aortic valve; IL-1β; Lazaroid U-74389G; NF-κB; Osteoblastic differentiation