title: Hyperbaric oxygen treatment prevents nitric oxide-induced apoptosis in articular cartilage injury via enhancement of the expression of heat shock protein 70.
abstract: Heat shock proteins (HSPs), inflammatory cytokines, nitric oxide (NO), and localized hypoxia-induced apoptosis are thought to be correlated to the degree of cartilage injury. We investigated the effect of hyperbaric oxygen (HBO) on (1) interleukin-1β (IL-1β)-induced NO production and apoptosis of rabbit chondrocytes and (2) healing of articular cartilage defects. For the in vitro study, RT-PCR and Western blotting were performed to detect mRNA and protein expressions of HSP70, inducible NO synthase (iNOS), and caspase 3 in IL-1β-treated chondrocytes. To clarify that the HSP70 was necessary for anti-iNOS and anti-apoptotic activity by HBO, we treated the cells with an HSP70 inhibitor, KNK437. For the in vivo study, cartilage defects were created in rabbits. The HBO group was exposed to 100% oxygen at 2.5 ATA for 1.5 h a day for 10 weeks. The control group was exposed to normal air. After sacrifice, specimen sections were sent for examination using a scoring system. Immunohistochemical analyses were performed to detect the expressions of iNOS, HSP70, and caspase 3. Our results suggested that HBO upregulated the mRNA and protein expressions of HSP70 and suppressed those of iNOS and caspase 3 in chondrocytes. KNK437 inhibited the HBO-induced downregulation of iNOS and casapase 3 activities. The histological scores showed that HBO markedly enhanced cartilage repair. Immunohistostaining showed that HBO enhanced HSP70 expression and suppressed iNOS and caspase 3 expressions in chondrocytes. Accordingly, HBO treatment prevents NO-induced apoptosis in articular cartilage injury via enhancement of the expression of heat shock protein 70.
authors.affiliation: Department of Orthopaedic Surgery and Hyperbaric Oxygen Therapy Center, Chang Gung Memorial Hospital, Kweishan, Taoyuan, Taiwan. wenneng@adm.cgmh.org.tw
authors.forename: Steve W N
authors.initials: SW
authors.lastname: Ueng
authors.forename: Li-Jen
authors.initials: LJ
authors.lastname: Yuan
authors.forename: Song-Shu
authors.initials: SS
authors.lastname: Lin
authors.forename: Chi-Chien
authors.initials: CC
authors.lastname: Niu
authors.forename: Yi-Sheng
authors.initials: YS
authors.lastname: Chan
authors.forename: I-Chun
authors.initials: IC
authors.lastname: Wang
authors.forename: Chuen-Yung
authors.initials: CY
authors.lastname: Yang
authors.forename: Wen-Jer
authors.initials: WJ
authors.lastname: Chen
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002358:Cartilage, Articular / Q000166:cytology / Q000293:injuries* / Q000502:physiology; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology* / Q000502:physiology*; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism; D006931:Hyperbaric Oxygenation / Q000379:methods*; D053583:Interleukin-1beta / Q000494:pharmacology; D009569:Nitric Oxide / Q000502:physiology*; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits
chemical_list: D018840:HSP70 Heat-Shock Proteins; D053583:Interleukin-1beta; D012333:RNA, Messenger; D009569:Nitric Oxide; D052247:Nitric Oxide Synthase Type II; D053148:Caspase 3; D010100:Oxygen