Effect Of Massage On TLR4/MyD88 Signal Transduction Pathway In Rat Knee Osteoarthritis Model

ObjectiveIn this study,we established a group of SPF-class SD female rats with knee osteoarthritis model by massage therapy,and tested various indicators of blood and knee cartilage after treatment:①serum IL-1β detected by ELISA,IL-6,TNF-α expression level;②knee joint cartilage Mankin score;③detection of TLR4,MyD88,TRAF-6,NF-κB p65 mRNA and protein expression in articular cartilage by real-time PCR and Western blot Level,through the statistical analysis of the test results,to explore the mechanism of massage therapy for knee osteoarthritis in rats,and provide a solid and reliable experimental basis for clinical treatment of knee osteoarthritis.MethodsFollowing the principle of randomization,10 rats were selected as blank group by random number table,and the remaining 40 rats were established by papain joint injection method.After successful modeling,the model was established.Rats were randomly divided into model group,massage group,gavage group,massage plus gavage group,10 rats in each group.The blank group and the model group were routinely raised,and the massage group was treated with a finger zen method.The gavage group was intragastrically administered with celecoxib solution,and the massage group was given the same two treatments.After 4 weeks of treatment,the serum levels of IL-1β,IL-6 and TNF-α were detected by ELISA in rats.Mankin score,real-time PCR,and cartilage detection by Western blot were performed on rat knee joint cartilage.TLR4,MyD88,TRAF-6,NF-κB p65 mRNA and protein expression levels.Statistics and processing data were measured using SPSS 19.0 software.Results①serum ELISA test results showed that the expression of serum IL-1β,IL-6 and TNF-α was up-regulated in each treatment group compared with the blank group,the difference was statistically significant(P<0.05);serum IL compared with the model group The expression of-1β,IL-6 and TNF-a was down-regulated,the difference was statistically significant(P<0.05);and the expression of serum IL-1β,IL-6 and TNF-α in each treatment group:massage plus gavage group<Tuina The difference between the group and the gavage group was statistically significant(P<0.05).②Mankin scores of knee articular cartilage showed that the scores of the treatment groups were higher than those of the blank group,the difference was statistically significant(P<0.05);the scores were lower than the model group,the difference was statistically significant(P<0.05);The cartilage Mankin scores of each treatment group:massage plus gavage group<Tuina group<gavage group,the difference was statistically significant(P<0.05).③The expression of TLR4 mRNA,MYD88 mRNA and NF-K B p65 mRNA in cartilage fluorescent quantitative PCR showed that the expression of TLR4 mRNA,γD88 mRNA and NF-κB p65 mRNA was up-regulated in the massage group and the gavage group(P<0.05).There was almost no difference between the massage group and the gavage group.(P>0.05);compared with the model group,the expression of each treatment group was down-regulated(P<0.05);the results of TRAF6 mRNA showed that the expression of each treatment group was up-regul.ated(P<0.05),and the expression of the model group was down-regulated(P<0.05);and the expression level between the treatment groups:massage plus gavage group<Tuina group<gavage group,the difference was statistically significant(P<0.05).④Western blot analysis of cartilage TLR4,MYD88,NF-KB p65 and TRAF6 protein showed that the expression of each treatment group was up-regulated compared with the blank group(P<0.05);the expression was down-regulated compared with the model group,the difference was statistically significant.Significance(P<0.05);and the expression between the treatment groups:massage plus gavage group<Tuina group<gavage group,the difference was statistically significant(P<0.05).ConclusionAccording to the experimental results,it can be seen that the massage therapy can down-regulate the expression of TLR4,MyD88,TRAF-6,NF-κB p65 mRNA and protein,and hinder the inflammatory reaction through the signal transduction pathway of TLR4/MyD88 signal transduction pathway.It inhibits the expression of downstream proinflammatory cytokines IL-1β,IL-6 and TNF-a,and plays a role in reducing inflammation and relieving pain and repairing articular cartilage,and finally achieves the purpose of preventing and treating knee osteoarthritis.This provides a basis for the study of the mechanism of action of the TLR4/MyD88 signal transduction pathway to prevent and treat knee osteoarthritis.