Study On Roles Of Diagnose And Traditional Chinese Medicine Treatment In OA Through Researching Levels Of MMP-3, MMP-9, NF-κB RANKL And NO In Peripheral Blood Of Knee OA Patients

Knee osteoarthritis (OA) is the most prevalent joint disease in humans accompaniedby pain, disability and joint deformities. The aim of our study was to research moresensitive diagnostic methods and available therapeutic tools in early stage of OA, throughexamining clinical and biochemical features of OA and investigating whether there is acorrelation between them. We would find therapeutic effects of shao ling plaster fordetumescence, through observing the effect of shao ling plaster for detumescence onplasma levels and transcript levels of correlation factors in knee osteoarthritis(OA)patients. This study included two parts.Part one: Objective Aim of this study was to judge whether there is a correlationbetween some biochemical features of knee osteoarthritic blood and clinical characteristicsand to evaluate the potential relationship between OA severity and putative biomarkers forthe disease. Methods Except heathy group(n=21), 105 patients suffering from knee OAwere analyzed clinically (Lequesne's index) and radiographically (Kellgren and Lawrence,K&L).Plasma and peripheral blood mononuclear cells (PBMC) were separately harvested.Specimens were analyzed concentrations of nitric oxide (NO), matrix metalloproteinase-3(MMP-3), matrix metalloproteinase-9 (MMP-9) and transcript levels of the receptoractivator of NF-kB ligand (RANKL) mRNA, MMP-3mRNA and MMP-9mRNA weremeasured using real-time quantitative RT-PCR. Results Data certified significantlyincreasing concentration of plasma MMP-3, MMP-9 and NO as well as transcript levels ofRANKLmRNA and MMP-9mRNA in the early OA (at gradeâ… ). There was a positivecorrelation of MMP-3, MMP-9 content in plasma and MMP-9mRNA expression levels inPBMC with the severity of clinical symptoms (total Lequesne's scores) in the early OA.NO content in plasma correlated with total Lequense's scores, paining scores at pressure,and swelling scores from the early OA patients (at gradeâ… ). Analogously, there werepositive correlations of RANKLmRNA expression respectively with total Lequense'sscores, paining scores at pressure, and swelling scores in OA patients at gradeâ… .Conclusion Results suggested that some biochemical factors, including content of NO, MMP-3, MMP-9 and transcript levels of some genes, including MMP-9mRNA andRANKLmRNA, may be specific and sensitive to diagnose OA diseases at early stage inthe pathological process of OA when radiological features do not reflect degradation ofarticular cartilage. Therefore, proper regulation of these factors and genes expressionlevels may be a promising and realistic new target for the treatment of degenerativeosteoarticular diseases.Part two: Objective To observe the effect of shao ling piaster for detumescence onplasma levels of MMP-3, MMP-9, NO and levels of MMP-3 mRNA, MMP-9 mRNA andRANKL mRNA in PBMC in knee OA patients at various grades. Methods Exceptheathy group(n=21), we did a randomised, double-blind placebo controlled trial, in which105 outpatients with knee osteoarthritis were randomly assigned shao ling plaster fordetumescence or placebo once daily for 2 weeks. Thirty-five patients wereradiographically classified as grade 1, 31 as grade 2, 20 as grade 3, and 19 as grade 4according to a modified version of the Kellgren and Lawrence scale. Plasma and PBMCwere obtained from all people. In both prior-treatment and post-treatment, we measuredplasma levels of MMP-3, MMP-9, NO and levels of MMP-3 mRNA, MMP-9 mRNA andRANKL mRNA in PBMC. Symptoms were also scored by Lequesne's index includingknee pain at rest, at movement, and at pressure, knee swelling, morning stiffness andwalking ability. Results Compared with findings in the prior-treatment, values for theconcentrations of plasma NO, MMP-3 and total Lequense scores in the post-treatment,were significantly lower in the grade 1 and 2 patients. It is the same to levels of PBMCMMP-9mRNA, RANKLmRNA and Lequesne scores in grade 1, and scores of knee painat pressure and knee swelling in grade 1, 2, and 4. In shao ling plaster for detumescencegroup, linear logistic regression analysis showed a signifcant postive correlation betweenconcentrations of serum MMP-3 and Lequesne scores in the grade 1 and 2 patients,whereas no correlation between levels of MMP-3mRNA and Lequesne scores. It was alsoshowed that a signifcant postive correlation between levels of PBMC RANKLmRNA andLequesne scores in grade 1 patients, and between levels of PBMC RANKLmRNA andscores of knee pain at pressure and knee swelling in grade 1, 2, and 4. Conclusion Thesefindings made us think that during shao ling plaster for detumescence therapy on kneeostearthiris, one of action mechanism was decreased concentrations of serum MMP-3 by down regulation of MMP-3mRNA expression in synoviocytes and chondrocytes ofarticular cartilage, sequently amended knee joints failure and improved functions of kneejoints in grade land grade 2; Decreasing the concentration of serum NO and inhibiting theexpression of MMP-9 mRNA in PBMC by various ways might also be the actionmechanisms of shao ling plaster for detumescence, so the clinical symptom ofosteoarthritis is improved and relieved by inhibiting the apoptosis of chondrocytes and thedegradation of cartilage matrix ; another was down regulation expression levels of PBMCRANKLmRNA to inhibite osteoclasts' action, to finally achieved the aim for treatmentOA patients in grade 1 and active stage.