Peroxynitrite induces gene expression in intervertebral disc cells. Export

@article{citeulike:5887670, abstract = {STUDY DESIGN: In vitro stimulation of human intervertebral disc (IVD) cells. OBJECTIVE: To investigate the oxidative/nitrosative effects of peroxynitrite on human nucleus pulposus (NP) cells. SUMMARY OF BACKGROUND DATA: Peroxynitrite is an important tissue-damaging species generated at sites of inflammation and degeneration. The aim of this study was to examine the effects of oxidative/nitrosative stress caused by peroxynitrite and the peroxynitrite donor SIN-1 in human NP cells. METHODS: Degenerated human IVD tissue was analyzed for nitrosylation by immunofluorescence. In addition, human NP cells were isolated from IVDs, expanded and stimulated either with peroxynitrite itself or a stable peroxynitrite donor (SIN-1). Nitrosylation, accumulation of intracellular reactive oxygen species, NF-kappaB nuclear translocation, and cell viability were analyzed by fluorescence. Gene expression of TNF-alpha, IL-1beta, IL-6, IL-8, and IL-10 was quantified by real-time (RT)-PCR. RESULTS: Degenerated IVD tissue showed strong nitrosylation, especially in the NP. Isolated human NP cells showed a strong signal for nitrosylation and intracellular reactive oxygen species on stimulation with peroxynitrite or SIN-1. NF-kappaB/p65 sustained nuclear translocation of NF-kappaB/p65 and stimulation of IL-1beta, IL-6, and IL-8 expression was noted on treatment of cells with SIN-1. CONCLUSION: This study provides evidence that peroxynitrite may play a role in disc degeneration and discogenic back pain development by an increased synthesis of proinflammatory cytokines. Nuclear translocation of NF-kappaB was identified as the potential underlying pathway. Therefore, neutralizing peroxynitrite and its derivatives (e.g., via the use of antioxidants) may be a novel treatment option for discogenic back pain.}, author = {Poveda, Lucy and Hottiger, Michael and Boos, Norbert and Wuertz, Karin}, citeulike-article-id = {5887670}, citeulike-linkout-0 = {http://dx.doi.org/10.1097/BRS.0b013e31819f2330}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/19407676}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=19407676}, day = {15}, doi = {10.1097/BRS.0b013e31819f2330}, issn = {1528-1159}, journal = {Spine}, keywords = {antioxidants, disc, nutrition}, month = {May}, number = {11}, pages = {1127--1133}, posted-at = {2009-10-04 20:54:43}, priority = {2}, title = {Peroxynitrite induces gene expression in intervertebral disc cells.}, url = {http://dx.doi.org/10.1097/BRS.0b013e31819f2330}, volume = {34}, year = {2009} }

TY - JOUR ID - citeulike:5887670 L3 - citeulike-article-id:5887670 N2 - STUDY DESIGN: In vitro stimulation of human intervertebral disc (IVD) cells. OBJECTIVE: To investigate the oxidative/nitrosative effects of peroxynitrite on human nucleus pulposus (NP) cells. SUMMARY OF BACKGROUND DATA: Peroxynitrite is an important tissue-damaging species generated at sites of inflammation and degeneration. The aim of this study was to examine the effects of oxidative/nitrosative stress caused by peroxynitrite and the peroxynitrite donor SIN-1 in human NP cells. METHODS: Degenerated human IVD tissue was analyzed for nitrosylation by immunofluorescence. In addition, human NP cells were isolated from IVDs, expanded and stimulated either with peroxynitrite itself or a stable peroxynitrite donor (SIN-1). Nitrosylation, accumulation of intracellular reactive oxygen species, NF-kappaB nuclear translocation, and cell viability were analyzed by fluorescence. Gene expression of TNF-alpha, IL-1beta, IL-6, IL-8, and IL-10 was quantified by real-time (RT)-PCR. RESULTS: Degenerated IVD tissue showed strong nitrosylation, especially in the NP. Isolated human NP cells showed a strong signal for nitrosylation and intracellular reactive oxygen species on stimulation with peroxynitrite or SIN-1. NF-kappaB/p65 sustained nuclear translocation of NF-kappaB/p65 and stimulation of IL-1beta, IL-6, and IL-8 expression was noted on treatment of cells with SIN-1. CONCLUSION: This study provides evidence that peroxynitrite may play a role in disc degeneration and discogenic back pain development by an increased synthesis of proinflammatory cytokines. Nuclear translocation of NF-kappaB was identified as the potential underlying pathway. Therefore, neutralizing peroxynitrite and its derivatives (e.g., via the use of antioxidants) may be a novel treatment option for discogenic back pain. IS - 11 JF - Spine SN - 1528-1159 EP - 1133 TI - Peroxynitrite induces gene expression in intervertebral disc cells. VL - 34 SP - 1127 KW - antioxidants KW - disc KW - nutrition AU - Poveda, Lucy AU - Hottiger, Michael AU - Boos, Norbert AU - Wuertz, Karin PY - 2009/05/15/ UR - http://dx.doi.org/10.1097/BRS.0b013e31819f2330 ER -