| Purpose : To observe the effects of electroacupuncture through needling on cartilage degeneration,TLR4 / My D88 / NF-κB signal pathway and serum related inflammatory factors in knee osteoarthritis model rats,and to explore the mechanism of electroacupuncture through needling on synovitis in knee osteoarthritis rats,so as to provide a substantial theoretical basis for clinical treatment and further research.Material and method:(1)Sixty male SD rats of SPF grade were randomly divided into sham operation group,model group,electroacupuncture penetration group and ordinary electroacupuncture group,with 15 rats in each group.The model was made after adaptive feeding.The sham operation group only opened the joint cavity of the right hind limb without any operation,that is,suture.The rats in other groups were made by anterior cruciate ligament disconnection.The rats were forced to exercise after operation,and the model was established for 8 weeks.(2)The sham operation group and model group did not receive intervention treatment,but only routine feeding.In the electroacupuncture penetration group,Dubi and Neixiyan were penetrated by 5-8mm,and the Xuehai and Liangqiu were penetrated by 5-8mm.Dubi and Neixiyan are connected to a group of circuits,and the Xuehai and Liangqiu are connected to a group of circuits.The density wave is selected,with the frequency of 2 Hz / 10 Hz and the intensity of 0.5-1.5 m A,once a day,20 minutes each time,10 days as a course of treatment.The interval between courses of treatment is 2 days,a total of 2 courses of treatment.In the ordinary electroacupuncture group,Dubi and Neixiyan were pricked obliquely at an angle of 30 ° to the skin for 3-5mm,Xuehai and Liangqiu were pricked straight for 3-5mm.Except for the pricking method,other parameters were the same as those in the electroacupuncture penetration group.(3)Muscle bone ultrasound was used to observe the effusion of knee joint in rats;the synovium of rats was stained with he to observe the synovium,and the cartilage of rats was stained with he,toluidine blue and Masson.The Mankin score was calculated to evaluate the cartilage degeneration;the relative expression contents of TLR4,My D88,TRAF-6 and NF-κB p65 protein in rat synovium were measured by Western blot,the expression of NF-κB p65 in rat synovium was measured by immunohistochemistry,and the contents of IL-1β,IL-6 and TNF-α in rat serum were measured by ELISA.Results:(1)Ultrasound showed that the area of liquid dark area in the sham operation group was smaller than that in the other three groups;Compared with the model group,the area of liquid dark area in electroacupuncture penetration group and ordinary electroacupuncture group was significantly reduced;And the reduction in the electroacupuncture penetration group was more obvious than that in the ordinary electroacupuncture group.(2)The Mankin score of rat cartilage showed that the score of model group was significantly higher than that of sham operation group(P<0.01);Compared with the model group,the scores of electroacupuncture through acupuncture group and ordinary acupuncture group were significantly lower(P<0.01).(3)The relative expression of TLR4,My D88,TRAF-6 and NF-κB p65 protein in rat synovium was measured by Western blot.Compared with the sham operation group,the expression levels of TLR4,My D88,TRAF-6 and NF-κB p65 protein in the synovium of the model group were significantly higher(P<0.01);Compared with the model group,the expression levels of TLR4,My D88,TRAF-6 and NF-κB p65 protein in synovium of electroacupuncture penetration group and ordinary electroacupuncture group were significantly lower(P<0.01);Compared with the electroacupuncture group,the expression levels of TLR4,My D88,NF-κB p65 protein and TRAF-6 protein in the ordinary electroacupuncture group increased(P<0.01),but the difference was not statistically significant(P>0.05).(4)The expression of NF-κB p65 protein in synovium of model group was significantly higher than that of sham operation group(P<0.01);Compared with the model group,the expression level of NF-κB p65 protein in synovium of electroacupuncture penetration group and ordinary electroacupuncture group decreased significantly(P<0.01);Compared with the electroacupuncture group,the expression level of NF-κB p65 protein in the ordinary electroacupuncture group increased,but the difference was not statistically significant(P>0.05).(5)The levels of serum IL-1β,IL-6 and TNF-α in the model group were significantly higher than those in the sham operation group(P<0.01);After treatment,compared with the model group,the contents of serum IL-1β,IL-6 and TNF-α in electroacupuncture penetration group and ordinary electroacupuncture group decreased significantly(P<0.01);IL-6 and TNF-α in electroacupuncture penetration group.The content of IL-1β was significantly lower than that of ordinary electroacupuncture group(P<0.01),and the content of IL-1β was lower than that of ordinary electroacupuncture group(P<0.05).Conclusion : Electroacupuncture through needling can significantly improve the morphological manifestations of cartilage and synovium of knee joint in koa rats,reduce the Mankin score of cartilage and reduce knee joint effusion;down regulate the expression of TLR4,My D88,TRAF-6 and NF-κB p65 protein in synovium of KOA rats and down the expression of IL-1β,IL-6,TNF-α in serum.Its mechanism is to inhibit the downstream inflammatory cascade by blocking the transduction of TLR4 / My D88 / NF-κB signal pathway,so as to improve knee synovitis. |
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