ECTS Abstracts (2015) 1 P272

NO Production by Sodium Nitroprusside in Cruciate Ligament Fibroblasts: the Associated Mechanisms

Chih-Min Kuo1 & Herng-Sheng Lee2

1Graduate Institute of Pathology and Parasitology, National Defense Medical Centre, Taiwan ROC, Taiwan; 2Department of Pathology and Laboratory Medicine, Kaohsiung Veterans General Hospital, Taiwan ROC, Taiwan.

Cruciate ligaments (CL) stabilise the joint structure while allowing a very large range of motion. CL injuries in particular are known to lead to development of early osteoarthritis (OA). Oxidative stress and inflammatory reaction may mediate muscle dysfunction by CL injuries. This study is designed to examine the oxidative stress and the modulation in fibroblasts isolated from CL. Sodium nitroprusside (SNP) was used as a NO inducer. The primary cultures of fibroblasts from knee joint CL tissues were treated with SNP at concentrations of 0, 0.1, 0.2, 0.3, 0.4 and 0.5 mM for 24 hours. Measurement of NO was done by Griess reaction. Gene expression and regulation was validated by real-time PCR. Exposure of SNP to fibroblasts showed significant increase of NO production in a dose dependent manner. At the same time, upregulation of iNOS, MMP1 and MMP13, but not MMP2, 3, and 9, was identified. The signal transduction involved the phosphorylation of NF-kappaB transcriptional factor. The selective inhibitor of NF-kappaB, parthenolide, showed inhibition of NO production. Further studies to investigate the antioxidants will be carried out.

Disclosure: The authors declared no competing interests.

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