Clinical Effect Of Meridian-Sinew Release Therapy On Moderate Knee Osteoarthritis And Its Effect On TLR4/MYD88/NF-?B Signaling Transduction Pathway

Objectives:1.To explore the clinical superiority of minimally invasive meridian therapy in early to mid-term knee osteoarthritis(KOA)through retrospective analysis of the short-and long-term therapeutic effects of minimally invasive meridian therapy in 108 patients with early to mid-term KOA.2.To verify the clinical efficiency of minimally invasive meridian therapy in the treatment of KOA at the molecular and gene expression level by investigating the TLR4/MYD88/NF-?B signaling pathway in synovial tissue of KOA patients before and 12 weeks after treatment.Methods:1.Patients who were consistent with American College of Rheumatology(ACR)KOA diagnosis standard and had grade I-III KOA based on the Kellgren-Lawrence radiographic classification system were selected,and from which 108 early to mid-term KOA patients(163 knees)were qualified according to the inclusion,exclusion,elimination,and drop-out criteria and underwent minimally invasive meridian therapy.These patients were followed-up.Therapeutic efficiency was evaluated using the index score of osteoarthritis(ISOA)1 month,1,2,3,4,and 5 years after treatment.2.Synovial tissue samples in joints were collected from the patients who were qualified and enrolled in the study according to the selection criteria during acupotomy before and 12 weeks after the minimally invasive meridian therapy,and the following experiments were conducted:(1)HE staining of the tissue to observe synovial hyperplasia,inflammatory infiltration and interstitial fibrosis;(2)RT-PCR to detect the gene expression of IL-1?,IL-6,IL-8,and MMP-13,Western blot to detect the protein level of TLR4/MYD88/NF-?B,and immunohistochemical assay to determine the protein expression of TLR4/MyD88/NF-?B/MMP-13 in synovial tissue.The experimental results were statistically analyzed.Results:1.Clinical study:The scores of pain,stiffness,and function as well as the WOMAC scores of early to mid-term KOA patients after minimally invasive meridian therapy were decreased compared with that before treatment;and the overall effective rate 1 month,1,2,3,4 and 5 years after treatment were 98.9%,96.7%,95.6%,87.9%,83.5%,and 78.0%,respectively.2.Experimental study:(1)Synovial hyperplasia,inflammatory infiltration and interstitial fibrosis after the minimally invasive meridian therapy were significantly reduced compared with that before treatment;(2)Gene expression of inflammatory cytokines,including IL-1?,IL-6,IL-8,TNF-?,and MMP-13,in synovial tissue were increased to certain degrees after treatment compared with that before treatment,and the differences were statistically significant(p<0.05);protein level of TLR4,MyD88,p-NF-?B,and p-I?B-a in synovial tissue after treatment was significantly lower than that before treatment,and the differences between the two groups was statistically significant(p<0.05);no significant difference in the protein level of NF-?B and I?B-? before and after treatment was noted(p>0.05);immunohistochemistry results showed that the TLR4,MyD88,MMP-13,p65,and I? B-?were stained brownish yellow after reaction,and more cells were stained and densely distributed in synovial tissues before treatment;after the treatment,these proteins were stained light yellow in synovial tissues with less cells stained and sparsely distributed;semi-quantitative immunohistochemical analysis revealed that the expression of TLR4,MyD88,MMP-13,p65,and I?B-? proteins were decreased after treatment,and the difference between the two groups was statistically significant(p<0.05).Conclusions:1.Clinical study:the long-term curative effect of minimally invasive meridian treatment in early and mid-term KOA is definite and the follow-up with 5-year outcome as endpoint showed that the treatment delays the progression of KOA with a favorable long-term effect.2.Experimental study:minimally invasive meridian therapy can effectively relieve the inflammatory infiltration,synovial hyperplasia,and interstitial fibrosis of synovial tissue,reduce the gene expression of IL-1(3,IL-6,IL-8,TNF-?,and MMP-13 as well as the protein level of TLR4/MyD88/p65/I?B-a/p-I?B-a in synovial tissue,and inhibit the expression of downstream molecules,such as MYD88 and NF-?B,via regulating the TLR4 signaling pathway,and block the secretion of inflammatory cytokines,so as to reduce the synovial inflammation,relieve symptoms,and delay the progression of KOA.Therefore,the definite therapeutic effect of minimally invasive meridian therapy on KOA is verified at the molecular and gene expression level.