Effects of nitric-oxide synthase inhibitor on matrix metalloproteinase's expression in osteoarthritls cartilage by Luminex analysis / 中华风湿病学杂志

ABSTRACT

Objective To investigate the effects of nitric-oxide synthase inhibitor on matrix metalloproteinases (MMPs) expression in osteoarthritis (OA) cartilage by Luminex analysis,and to explore the mechanism of nitric-oxide synthase inhibitor on modification of the metabolism of OA cartilage.Methods Fifteen specimens of articular cartilage taken from the patients with OA were cultured and divided in two groups.The control group was those with no intervention.L-NIL group was co-cultured with NOS inhibitor L-NIL.After 72 h cultivation,the release of NO and the activity of NOS on OA cartilage were measured by Griess reaction and spectrophotometric methods.MMPs (MMP-1,MMP-2,MMP-3,MMP-9,MMP-13) expression was measured by Luminex analysis.Comparisons between groups were performed with paired sampies t test.Results After cultured for 72 h,spectrophotometric analysis showed high concentration of NO release[(216±47) μmol/L ] and high level of active NOS [(5.7±1.3)U/ml]in supernatants of the control,1 mmol/L concentration L-NIL could evidently reduce NO release [(55±20)μmol/L,P<0.01] and NOS activity [(1.7±0.7)U/ml,P<0.01 ].Luminex analysis demonstrated high MMP-1,MMP-2,MMP-3,MMP-9,MMP-13 expression in cartilages of the control group [respectively for (10.8±5.4)ng/ml,(9.2±3.3) ng/ml,[11.6±4.2 )ng/ml,(1.27±1.07)ng/ml,(3.6±1.3)ng/ml] and 1 mmol/L concentration L-NIL could evidently inhibit MMP-1,MMP-2,MMP-3,MMP-9,MMP-13 expression [respectively for (3.6±1.8)ng/ml,(2.3±1.2)ng/ml,(3.6±1.4)ng/ml,(0.65±0.21)ng/ml,(1.8+0.5)ng/ml,P<0.05 ].Conclusion Luminex analvsis has shown that NOS inhibitor can reduce NO release and NOS activity and modify metabolism of articular cartilage by inhibiting the over-expression of MMPs.