PMID- 26542776 OWN - NLM STAT- MEDLINE DCOM- 20160720 LR - 20220331 IS - 1478-6362 (Electronic) IS - 1478-6354 (Print) IS - 1478-6354 (Linking) VI - 17 DP - 2015 Nov 5 TI - Excessive reactive oxygen species are therapeutic targets for intervertebral disc degeneration. PG - 316 LID - 10.1186/s13075-015-0834-8 [doi] LID - 316 AB - INTRODUCTION: Oxidative stress has been reported to be involved in numerous human diseases, including musculoskeletal disorders such as osteoarthritis. However, the interaction between intervertebral disc (IVD) degeneration and oxidative stress is not well understood. The purpose of the present study was to elucidate the contribution of oxidative stress to IVD degeneration and the efficacy of antioxidant treatment for degenerative discs. METHODS: The expression level of an oxidative stress marker, nitrotyrosine, was assessed by immunohistochemistry and Western blotting. For evaluating intracellular reactive oxygen species (ROS) levels and oxidative stress in rat annulus fibrosus (AF) cells, flow cytometry and luciferase assay with an OKD48 construct were performed. The grade of IVD degeneration was assessed by magnetic resonance imaging and histological analysis. RESULTS: A high frequency of nitrotyrosine-positive cells was observed in rat and human degenerative discs. mRNA expression of catabolic factors such as tumor necrosis factor-alpha (TNF-alpha), matrix metalloprotease-3 (MMP-3), and cyclooxygenase-2 (COX-2) was significantly induced by treatment with H2O2 or buthionine sulfoximine, whereas that of aggrecan, an important chondrogenic proteoglycan, was reduced in a dose-dependent manner. Treatment with mitogen-activated protein kinase (MAPK) inhibitors blocked the inductive effect of excessive ROS on COX-2 mRNA expression. Western blotting confirmed the phosphorylation of MAPKs in H2O2 and BSO-treated AF cells. Conversely, we showed that TNF-alpha induced oxidative stress with increased intracellular ROS levels in AF cells. Treatment with the antioxidant N-acetyl cysteine (NAC) abrogated the catabolic effect of excessive ROS and TNF-alpha in vitro. Finally, we showed that oral administration of NAC prevented IVD degeneration in rat degenerative model. CONCLUSIONS: A positive feedback loop was formed between excessive ROS and TNF-alpha in AF cells. Thus, oxidative stress contributes to the progression of IVD degeneration and NAC can be a therapeutic option for IVD degeneration. FAU - Suzuki, Satoshi AU - Suzuki S AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. ssatosea@yahoo.co.jp. FAU - Fujita, Nobuyuki AU - Fujita N AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. nfujita2007@yahoo.co.jp. FAU - Hosogane, Naobumi AU - Hosogane N AD - Department of Orthopaedic Surgery, National Defense Medical College, 3-2 Namiki, Tokorozawa, Saitama, 359-8513, Japan. hosonao@outlook.com. FAU - Watanabe, Kota AU - Watanabe K AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. watakota@gmail.com. FAU - Ishii, Ken AU - Ishii K AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. keni8888@z7.keio.jp. FAU - Toyama, Yoshiaki AU - Toyama Y AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. toyama@z6.keio.jp. FAU - Takubo, Keiyo AU - Takubo K AD - Department of Stem Cell Biology, Research Institute, National Center for Global Health and Medicine, 1-21-1 Toyama, Shinjuku-ku, Tokyo, 162-8655, Japan. keiyot@gmail.com. FAU - Horiuchi, Keisuke AU - Horiuchi K AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. keisukehoriuchi@gmail.com. FAU - Miyamoto, Takeshi AU - Miyamoto T AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. miyamoto@z5.keio.jp. FAU - Nakamura, Masaya AU - Nakamura M AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. masa@a8.keio.jp. FAU - Matsumoto, Morio AU - Matsumoto M AD - Department of Orthopaedic Surgery, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo, 160-8582, Japan. morio@a5.keio.jp. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20151105 PL - England TA - Arthritis Res Ther JT - Arthritis research & therapy JID - 101154438 RN - 0 (Antioxidants) RN - 0 (Reactive Oxygen Species) RN - WYQ7N0BPYC (Acetylcysteine) SB - IM MH - Acetylcysteine/pharmacology MH - Adult MH - Aged MH - Animals MH - Antioxidants/*pharmacology MH - Blotting, Western MH - Female MH - Flow Cytometry MH - Humans MH - Intervertebral Disc/drug effects/metabolism MH - Intervertebral Disc Degeneration/*pathology MH - Magnetic Resonance Imaging MH - Male MH - Middle Aged MH - Rats MH - Rats, Wistar MH - Reactive Oxygen Species/*metabolism MH - Real-Time Polymerase Chain Reaction PMC - PMC4635526 EDAT- 2015/11/07 06:00 MHDA- 2016/07/21 06:00 PMCR- 2015/11/05 CRDT- 2015/11/07 06:00 PHST- 2015/04/26 00:00 [received] PHST- 2015/10/21 00:00 [accepted] PHST- 2015/11/07 06:00 [entrez] PHST- 2015/11/07 06:00 [pubmed] PHST- 2016/07/21 06:00 [medline] PHST- 2015/11/05 00:00 [pmc-release] AID - 10.1186/s13075-015-0834-8 [pii] AID - 834 [pii] AID - 10.1186/s13075-015-0834-8 [doi] PST - epublish SO - Arthritis Res Ther. 2015 Nov 5;17:316. doi: 10.1186/s13075-015-0834-8.