| Research backgroundWith the advent of an aging society,the incidence of knee osteoarthritis(KOA)is gradually increasing.It is particularly important for early intervention in patients with KOA to reduce the incidence of joint replacement surgery.Electroacupuncture has the advantages of good analgesic effect and easy operation in the treatment of KOA,but the mechanism of its treatment of KOA is not very clear.This study further explored the clinical efficacy and mechanism of electroacupuncture in treating KOA from two aspects:clinical research and basic research.Objective1.The acupoints were determined under the guidance of the theory of meridians,electroacupuncture was selected to intervene knee osteoarthritis,to observe the improvement of pain,functional activity and quality of life,as well as the measurement of muscle strength of flexor and extensor muscles around the knee joint,and to comprehensively analyze the therapeutic effect of electroacupuncture on knee osteoarthritis2.Electroacupuncture was selected to intervene in the rabbit KOA model,to observe the mRNA and protein expression of Toll-like receptor and its main components on the signaling pathway in the sampled tissues,and to explore the expression differences of electroacupuncture on Toll-like receptor and its main components on the signaling pathway in the synovium and cartilage of KOA at different stages.Research method1.Clinical research1.1 Treatment planForty patients with mild to moderate KOA were enrolled.Randomly divided into electroacupuncture group and glucosamine sulfate capsule group.Electroacupuncture group:selected acupoints of Nei xiyan,Du bi,Liang qiu,Xue hai,Yang lingquan,Yin lingquan and Ashi point.Conventional acupuncture of all points,after getting qi,select Nei xiyan and Du bi a group,Liang qiu and Xue hai a group,connected to the Yingdi brand electroacupuncture instrument,density wave,for 25 minutes.Treatment was given 3 times a week for 4 consecutive weeksControl group:oral glucosamine sulfate capsules were used for treatment,0.628g each time,3 times a day,for 4 weeks.1.2 Observation index and time nodeMain efficacy indicators:visual analog score(VAS score),knee osteoarthritis index score(WOMAC score);Isokinetic muscle strength test indicators:include peak moment(PT),total work done(TW)and average power(AP);Secondary efficacy indicators:quality of life health questionnaire score(sf-36)Evaluation point:three time nodes were recorded before treatment,2 weeks after treatment and 4 weeks after treatment2.Experiment research2.1 Experimental scheme44 rabbits were randomly divided into 6 groups,including 4 in blank group,8 in sham operation group,1 in model group,2 in model group,1 in electroacupuncture group and 2 in electroacupuncture group.The koa animal model was established by the improved Hulth method,and the experimental koa model rabbits were treated by electroacupuncture.Acupoint selection:Yanglingquan,yinlingquan,inner knee eye,calf nose,Liangqiu,Xuehai and other acupoints were treated with electroacupuncture for 20 minutes each time,3 times a week,for 4 weeks.After the intervention,samples were taken from model group 1 and electroacupuncture group 1 at 6 weeks after modeling,and from blank group,sham operation group,model group 2 and electroacupuncture group 2 at 8 weeks after modeling.The samples were taken from both knees in the blank group and left knee joints in the other groups2.2 Test indicatorsIn Experiment 1,the levels of IL-1? IL-6,TNF-?,MMP1,MMP3 and MMP13 in serum and joint fluid of each group were measured to observe the expression of cytokines in different groups of rabbits.He staining was used to observe the morphological changes of synovium and cartilage of experimental koa rabbits.In Experiment 2,the mRNA and protein expression of TLR4,TLR2,NF-?B,MyD88 and TRAF6 in synovium and cartilage samples of rabbits in each group were detected by real-time PCR and Western blot,and the results of each group were compared.Results1.1 Knee pain VAS scoreCompare within groups:The VAS score of the observation group was significantly lower than that of the control group(P<0.05)at 2 weeks and 4 weeks after treatment.Compared with the control group:the VAS score of each time node in the observation group was lower than that of the control group(P<0.05),indicating that electroacupuncture has a better analgesic effect.1.2 WOMAC index scoreCompare within groups:the difference between the two weeks after treatment and the four weeks after treatment was statistically significant(P<0.05);the difference between the four weeks after treatment and the two weeks after treatment was statistically significant(P<0.05).Compared with the control group:there was no significant difference between the two groups before treatment(P>0.05).The observation group was better than the control group in terms of joint pain,stiffness,body function and other dimensions.1.3 Sf-36 quality of life scale scoresPhysical health score:compared with the same time node control group,the scores of the two groups before treatment(P>0.05)were comparable,and the differences were statistically significant(P<0.05).The difference was statistically significant(P<0.05).The difference was statistically significant(P<0.05).Mental health score:compared with the same time node control group,the scores of the two groups before treatment(P>0.05)were comparable,and the differences were statistically significant(P<0.05).The difference was statistically significant(P<0.05).The difference was statistically significant(P<0.05).1.4 Isokinetic muscle force detectionComparison of PT value:Compare within groups:The PT values of flexor and extensor muscles in the electroacupuncture group were compared with those before treatment 2 weeks after treatment and 4 weeks after treatment respectively(P<0.05).Comparison between the control group after 2 weeks of treatment and before treatment(P<0.05),the difference between the control group after 4 weeks of treatment and before treatment was statistically significant(P<0.05)Compared with the control group:There was no significant difference between the two groups before treatment(P>0.05);the peak moment of extensor and flexor in the electroacupuncture group was better than that in the control group at the same time node(P<0.05);the difference between the two groups after treatment(P<0.05)the difference was statistically significantComparison of TW value:Compare within groups:The TW values of flexor and extensor muscles in the electroacupuncture group were compared with those before treatment 2 weeks after treatment and 4 weeks after treatment respectively(P<0.05).Comparison between the control group after 2 weeks of treatment and before treatment(P<0.05),the difference between the control group after 4 weeks of treatment and before treatment was statistically significant(P<0.05)Compared with the control group:The comparison of extensor group and flexor group muscle strength before treatment between the two groups was(P>0.05),which was comparable;the total work of extensor group and flexor group in the electroacupuncture group after 2 weeks and 4 weeks of treatment was better than that in the control group with the same time node(P<0.05),which was statistically significant.Comparison of AP value:Compare within groups:The AP values of flexor and extensor muscles in the electroacupuncture group were compared with those before treatment 2 weeks after treatment and 4 weeks after treatment respectively(P<0.05).Comparison between the control group after 2 weeks of treatment and before treatment(P<0.05),the difference between the control group after 4 weeks of treatment and before treatment was statistically significant(P<0.05)Compared with the control group:There was no statistical significance between the two groups before treatment(P>0.05);the average power of the extensor group and flexor group in the electroacupuncture group was better than that of the control group at the same time node(P<0.05);the muscle power of the extensor group and flexor group in the electroacupuncture group was improved to different degrees in the two weeks after treatment and the four weeks after treatment(P<0.05)2.Experiment researchExperiment 1ELISA results:compared with the normal group,there was no significant difference in the levels of IL-1 P,TNF-?,IL-6,MMP1,MMP3 and MMP13 metalloproteinases in the serum and joint fluid of the sham operated group koa rabbits(P>0.05).The levels of IL-1?,TNF-?,IL-6,MMP1,MMP3 and MMP13 in serum and joint fluid of the two model groups were significantly higher than those of the two electroacupuncture treatment groups and the sham operation group(P<0.05).Pathological changes of synovium:compared with the blank group and the sham operation group,the synovium tissue of the two model groups was congested,and the inflammatory infiltration of local lymph,monocyte and plasma cell was obvious;even synovium hyperplasia,fibrosis and capillary sclerosis were involved in the two model groups.The proliferation of synovium and infiltration of lymphocyte in electroacupuncture group 1 and electroacupuncture group 2 were significantly less than those in the two model groups.Pathological changes of articular cartilage:compared with the blank group and the sham operation group,the surface of cartilage in the two model groups was not smooth,the arrangement of cells was not regular,the integrity of tidal line was not good,and even superficial ulcer appeared in the two model groups,the thickness of cartilage became thinner,the tidal line moved forward and disordered;the contour was not clear,the arrangement of cells was not regular,the tidal line was slightly disordered,the stratification was not clear,but the overall table was not clear Now it is better than the model group.Experiment 2:Real Time PCR results:Comparison of mRNA expression levels of TLR4,NF-B,MyD88,TLR2 and TRAF6 in synovium:the mRNA expression level of model 1 and model 2 group was significantly higher than that of sham operation group and blank control group(P<0.05).The mRNA expression level of the two electroacupuncture groups was lower than that of the two model groups(P<0.05),and the difference was statistically significant.Comparison of mRNA expression levels of TLR4,NF-B,MyD88,TLR2 and TRAF6 in cartilage:The mRNA expression level of model 1 and model 2 group was significantly higher than that of sham operation group and blank control group(P<0.05).The mRNA expression level of the two electroacupuncture groups was lower than that of the two model groups(P<0.05),and the difference was statistically significant.Western Blot results:Protein expression of TLR4,NF-B,MyD88,TLR2 and TRAF6 in synovium::the protein expression of model 1 group and model 2 group was significantly higher than that of blank group and sham group(P<0.05).The ratio of protein expression level in the two electroacupuncture groups was significantly lower than that in the two model groups(P<0.05),and the difference was statistically significantProtein expression of TLR4,NF-B,MyD88,TLR2 and TRAF6 in cartilage:The protein expression of model 1 group and model 2 group was significantly higher than that of blank group and sham group(P<0.05).The ratio of protein expression level in the two electroacupuncture groups was significantly lower than that in the two model groups(P<0.05),and the difference was statistically significant.Conclusion1.Electroacupuncture in the treatment of knee osteoarthritis in the early and middle stage can effectively relieve joint pain,improve joint function,enhance the muscle strength of flexor and extensor muscles around the knee joint,and improve the quality of life of patients2.The levels of cytokines and matrix metalloproteinases were correlated with the severity of KOA..Electroacupuncture can effectively inhibit the levels of cytokines and matrix metalloproteinases in serum and joint fluid,which is helpful to control joint inflammation3.The expression of Toll like receptor is correlated with the severity of knee osteoarthritis.Electroacupuncture can effectively inhibit the mRNA and protein expression of TLRs/MyD88/NF-?B signal pathway related elements,suggesting that acupuncture may alleviate joint inflammation by inhibiting this inflammatory response signal pathway. |
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