| Background:Acupuncture has a good clinical effect in the treatment of rheumatoid arthritis,but its mechanism of action is still unclear.In this study,adjuvant arthritis rats were used to explore the molecular mechanism of electroacupuncture for anti-inflammatory,providing objective experimental basis for clinical efficacy.Objective:To observe the effect of electroacupuncture on the expression of mmp-3 and mmp-9 in adjuvant arthritis rats at “zusanli” and “xuan zhong”points,and to explore the molecular mechanism of electroacupuncture inhibiting the inflammatory response and synovial hyperplasia of AA rats.Methods: SD rats were randomly divided into control group,model group,acupoint group and non-acupoint group,with 10 rats in each group.The rat model of arthritis was prepared by injecting freund's complete adjuvant into the foot pad,and the control group was simulated with normal saline.Rats in the acupoint group received the electroacupuncture on both sides of“zusanli” and “xuan zhong” points,2Hz,15 min/ time,once every 2 days,a total of 8 times.The non-acupoint group was acupuncture through the skin at the point “zusanli” and “xuan zhong"”which was opened 5mm to the side,and the electric stimulation intensity was the same as that of the acupoint group.The control group and the model group did not intervene.The arthritis index score system was used to evaluate the arthritis index of model rats on day 3,day 8,day 13 and day 18,respectively.On the 18 th day,the rats were sacrificed,abdominal aortic blood and synovial tissue of the ankle joint were collected,and the pathomorphological changes of the ankle jointwere observed by HE staining.Tunnel method was used to detect the apoptosis of synovial cells in the ankle joint of model rats.MRNA expression of mmp-3and mmp-9 in synovium of ankle joint was detected by rt-pcr,and protein expression of mmp-3 and mmp-9 in synovium of ankle joint was detected by western blot.The expression of MMP-3 and MMP-9 in serum of rats in each group was detected by enzyme-linked immunosorbent assay(ELISA).Results:1.The results of arthritis index showed that at each time point,the joint index score of rats in the model group was significantly higher than that in the control group(P<0.05).On the 18 th day,the arthritis index score of rats in the acupoint group was significantly lower than that in the model group and the non-acupoint group(P<0.05).2.The pathological sections showed that the synovium cells of the ankle joint of the model group and the non-acupoint group showed excessive proliferation,and the inflammatory cells increased significantly.The degree of synovium cell proliferation in the acupoint group was significantly lower than that in the model group and the non-acupoint group,and the number of inflammatory cells was less.3.Tunnel test results showed that the apoptosis rate of synovial cells in model group,acupoint group and non-acupoint group was significantly higher than that in control group(P<0.05).The apoptosis rate of synovial cells in the acupoint group was significantly higher than that in the model group(P<0.05).There was no significant difference in the apoptosis rate of synovial cells between the non-acupoint group and the model group(P<0.05).4.The results of rt-pcr showed that the expression levels of mmp-3 and mmp-9mrna in the model group were significantly higher than those in the control group(P<0.05).The expression levels of mmp-3 and mmp-9 mrna in the acupoint group were significantly lower than those in the model group and the non-acupoint group(P<0.05).There was no significant difference between the expression levels of mmp-3 and mmp-9 mrna in the non-acupoint group and the model group(P < 0.05).5.Western blot results showed that the expression levels of mmp-3 and mmp-9proteins in the model group were significantly higher than those in the control group(P<0.05).The expression levels of mmp-3 and mmp-9 proteins in the acupoint group were significantly lower than those in the model group and the non-acupoint group(P<0.05).There was no significant difference between the expression levels of mmp-3 and mmp-9 in the non-acupoint group and the model group(P < 0.05).6.The results of enzyme-linked immunosorbent assay(ELISA)showed that the serum levels of MMP-3 and MMP-9 in the model group were significantly higher than those in the control group(P<0.05).The serum levels of MMP-3 and MMP-9in acupoint group and non-acupoint group were significantly lower than those in model group(P<0.05).The serum levels of MMP-3 and MMP-9 in non-acupoint group were significantly higher than those in acupoint group(P<0.01).Conclusion:1.Electroacupuncture at “zusanli” and “xuanzhong” points can reduce joint inflammation in rats with adjuvant arthritis,improve the rate of synovial cell apoptosis,promote tissue repair,and control the progress of the disease.2.Electroacupuncture at “zusanli” and “xuanzhong” point can regulate the expression levels of mmp-3 and mmp-9 in serum and local tissues,promote the apoptosis of synovial cells,and then control the joint inflammation of adjuvant arthritis rats. |
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