Header

UZH-Logo

Maintenance Infos

Peroxynitrite induces gene expression in intervertebral disc cells


Poveda, L; Hottiger, M; Boos, N; Wuertz, K (2009). Peroxynitrite induces gene expression in intervertebral disc cells. Spine, 34(11):1127-1133.

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.

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.

Statistics

Citations

20 citations in Web of Science®
19 citations in Scopus®
Google Scholar™

Altmetrics

Downloads

0 downloads since deposited on 28 Feb 2010
0 downloads since 12 months

Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Department of Molecular Mechanisms of Disease
07 Faculty of Science > Department of Molecular Mechanisms of Disease

04 Faculty of Medicine > Balgrist University Hospital, Swiss Spinal Cord Injury Center
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:May 2009
Deposited On:28 Feb 2010 14:21
Last Modified:05 Apr 2016 13:56
Publisher:Lippincott Wiliams & Wilkins
ISSN:0362-2436
Publisher DOI:https://doi.org/10.1097/BRS.0b013e31819f2330
PubMed ID:19407676

Download