| Purpose:1.Document Research: With the help of ‘Traditional Chinese Medicine inheritance computing platform system’(TCMICS V3.0).To study the prescription of acupoint for treating knee osteoarthritis by Electroacupuncture.In order to explore the characteristics and selection of acupuncture points in the treatment of Knee-osteoarthritis.2.Clinical Research: To observe the clinical efficacy of Electroacupuncture point-through therapy on the treatment of Knee-osteoarthritis and its influence on Synovial-membrane,to verify the clinical efficacy of Electroacupuncture point-through therapy on the treatment of Knee-osteoarthritis,and in order to provide intuitive quantitative basis for its clinical application.3.Fundamental research: Based on NLRP3/ Caspase-1 /IL-1β signaling pathway,To observe the mechanism of Electroacupuncture point-through therapy on synoviocyte pyroptosis in rats with Knee-osteoarthritis.Material and method:1.Based on the TCMICS V3.0 to research the rules of Electroacupuncture prescription on the treatment of Knee-osteoarthritis:Through the method of ‘literature search-professional search’in CNKI.The related literatures of ‘Electroacupuncture therapy’ and ‘Electroacupuncture point-through therapy’ on the treatment of Knee-osteoarthritis(Knee-Bi disease in TCM)in the recent 25 years(from 1996 to 2021)were searched.Develop a scientific retrieval strategy.Standardize the names of acupoints and terms of symptoms.Data management,statistics and analysis were conducted on the frequency of acupoints,frequency of acupoints of various syndromes,distribution of meridians and collaterals,distribution of acupoints on the body surface,distribution of specific acupoints and association rules of acupoints combination by the TCMICS(V3.0).2.Clinical effect of Electroacupuncture point-through therapy on Knee-osteoarthritis:A total of 106 patients with Knee-osteoarthritis from the Department of orthopedics department K2 of the First Affiliated Hospital of Liaoning University of Traditional Chinese Medicine were selected as the research objects.The time limit of the study was from May 2021 to November2021.The eligible patients were randomly divided into treatment group and comparative group by random number table method,with 53 patients in each group.The comparative group was treated with Electroacupuncture combined with ordinary acupuncture,and the treatment group was treated with Electroacupuncture combined with Electroacupuncture point-through therapy.Take 10 days as a course of treatment,and 2 days of rest between the courses.All participants included comparative and treatment groups both have 2 courses of treatment.The changes of the following clinical indicators were detected before,during and after treatment in 2 groups,such as: Knee effusion,Thickness of Synovial,changes in synovial blood flow,VAS score,WOMAC arthritis index score,KOOS outcome score and adverse reactions.3.To investigate the effect of Electroacupuncture point-through therapy on synovial pyroptosis in rats with knee osteoarthritis based on NLRP3/ Caspase-1 /IL-1β signaling pathway:Sixty SPF male rats were divided into sham operation group,model group,treatment group and comparative group by random number table method,with 15 rats in each group.KOA model was made by cutting the anterior cruciate ligament of right hind limb of rats.In the experimental group,the right hindlimb calf nose and inner knee eyes were punctured,and the Xuehai and Liangqiu were punctured 5-8mm and connected with electroacupuncture,20min/d,10 days as a course of treatment,the treatment interval was 2 days,a total of 2 courses of intervention.The comparative group was treated with 3-5mm direct puncture of Dubi,Neixiyan,Xuehai and Liangqiu.The method of electrode connection and treatment course were the same as with the experimental group.Sham operation group and model group did not receive intervention treatment.At the end of the experiment,HE staining was used to observe the morphology of the synovium of the knee,musculoskeletal ultrasound was used to examine the fluid accumulation in the suprapatellar sac,ELISA was used to determine the expressions of IL-1β and IL-18 in serum,and Immunohistochemical method was used to measure the expressions of NLRP3 and GSDMD.Western Blot was used to determine the expression levels of NLRP3,GSDMD,Pro-Caspase-1,Pro-IL-1β,Caspase-1 and ASC proteins in synovial membrane of rats.Results:1.Based on the TCMICS V3.0 to research the rules of Electroacupuncture prescription on the treatment of Knee-osteoarthritis:According to the literature retrieval rules of CNKI,a total of834 literatures were retrieved,and a total of 514 literatures were included after screening,involving 86 acupoints with a total frequency of 3897 times.The frequency of ‘Dubi’ was the highest,reaching 484 times,accounting for 94.16%.There were 9 acupoints that appeared more than 100 times.The meridian with the largest number of acupoints is the bladder meridian,with 18 acupoints,accounting for 20.93%.The most frequent meridians were Stomach meridian of foot-yangming,reaching 1264 times,accounting for 32.44%.There were27 acupuncture prescriptions with definite syndrome type.In the aspect of acupoint selection,proximal acupoint selection accounted for 45 acupoints,accounting for 52.33%.The total frequency was 3,689 times,accounting for 94.66%.2.Clinical effect of Electroacupuncture point-through therapy on Knee-osteoarthritis:(1)There was no significant difference between the two groups in gender,age,weight,BMI index,disease site and other demographic data and clinical characteristics(P > 0.05).(2)Basic data of 106 patients with Knee-osteoarthritis were analyzed,including 74 female patients and 32 male patients.There were 4 patients with low BMI,39 patients with normal BMI,and 63 patients with high BMI.(3)In terms of synovial thickness of knee joint.Comparison within group A: comparison between before and after treatment,middle and after treatment in comparative group(P <0.05),comparison between before and after treatment(P > 0.05),comparison between before and after treatment,between before and after treatment,and between and after treatment in treatment group(P < 0.05).Comparison between group B: comparison between before treatment and middle group(P > 0.05),comparison between middle group after treatment(P <0.05).Knee joint effusion,comparison within group A: comparison between before and after treatment,middle and after treatment in comparative group(P < 0.05),comparison between before treatment and middle treatment in comparative group(P > 0.05).Comparison of treatment group before and after treatment,before and medium,before and medium(P <0.05).Comparison between group B: comparison between groups before treatment(P > 0.05),comparison between groups during and after treatment(P < 0.05).Articular synovial blood flow,comparison within group A: comparison between the comparative group before and after treatment,during and after treatment(P < 0.05),comparison between the comparative group before and after treatment(P > 0.05),comparison between the treatment group before and after treatment,during and after treatment(P < 0.05),comparison between the comparative group before and after treatment(P < 0.05).Comparison between group B:comparison between groups before treatment(P > 0.05),comparison between groups during and after treatment(P < 0.05).(4)In terms of comparison of VAS scores.Comparison within group A: comparison between comparative group before and after treatment,middle and after treatment(P < 0.05),comparison between comparative group during and after treatment(P > 0.05).Comparison between before and after treatment,between before and after treatment,between and after treatment(P < 0.05).Comparison between group B: the comparison between before and after treatment was poor(P > 0.05),and the comparison between after treatment was poor(P< 0.05).(5)In terms of comparison of WOMAC scores.Comparison within group A: comparison between before and after treatment,between before and middle,between and after treatment in comparative group(P < 0.05),comparison between before and after treatment,between before and middle,between and after treatment in treatment group(P < 0.05).Comparison between group B: comparison between before treatment and middle group(P > 0.05),comparison between middle group after treatment(P < 0.05).(6)In terms of comparison of KOOS scores:Comparison within group A: comparison between before and after,between before and middle,between and after in the comparative group(P <0.05),comparison between before and after,between before and middle,between and after in the treatment group(P < 0.05),comparison between before and middle in the treatment group(P > 0.05),comparison between between the treatment group and middle in the treatment group(P < 0.05).3.To investigate the effect of Electroacupuncture point-through therapy on synovial pyroptosis in rats with knee osteoarthritis based on NLRP3/ Caspase-1 /IL-1β signaling pathway:(1)In terms of Mankin’s score of knee cartilage of rats in each group: Compared with Mankin’s score of model group,treatment group and comparative group,the cartilage condition of rats in sham operation group was good,and the score decreased significantly(P < 0.05).Compared with model group,Mankin’s score in treatment group and comparative group was significantly decreased(P < 0.05).Compared with the treatment group,mankin’s score in the comparative group was increased(P < 0.05).(2)In terms of HE staining of the synovial membrane of the knee of rats in each group: the synovial structure of the sham operation group was normal;In model group,the number of synovial lining cells increased,collagen fiber tissue hyperplasia,a large number of lymphocytes,macrophages,neutrophils and other inflammatory cells infiltration,multiple angiogenesis.The number of cell layers in the lining layer of synovial membrane in the treatment group and the comparative group was less,the tissue area of collagen fiber was smaller,and inflammatory cell infiltration was less.(3)In terms of Ultrasonic examination of suprapatellar bursa effusion of rats in each group:only a very small amount of effusion was observed in the sham operation group;Compared with the sham operation group,the amount of effusion in the model group,the treatment group and the comparative group increased significantly.Compared with the model group,the amount of effusion in the treatment group was significantly less,approaching the normal synovial membrane.The amount of effusion in the comparative group decreased compared with the model group,but it was still significantly more than that in the treatment group.(4)In terms of the expression levels of IL-1β and IL-18 in serum of rats in each group showed that compared with sham operation group,the contents of IL-1β and IL-18 in serum of model group were significantly increased(P < 0.05).Compared with model group,the contents of SERUM IL-1β and IL-18 in treatment group and comparative group were significantly decreased(P < 0.05).Compared with the treatment group,the contents of IL-1β and IL-18 in the comparative group were increased(P < 0.05).(5)In terms of Immunohistochemical detection of NLRP3 and GSDMD expression:immunohistochemical analysis can be qualitative and quantitative analysis of substances to be tested.The expression of NLRP3 and GSDMD in synovial tissues of rats in each group was detected,and the contents of NLRP3 and GSDMD in synovial tissues of rats in model group were significantly increased compared with that in sham group(P < 0.05).After treatment,the contents of NLRP3 and GSDMD in synovial membrane of treatment group and comparative group were significantly decreased compared with model group(P < 0.05).In addition,compared with the treatment group,the contents of NLRP3 and GSDMD in the comparative group were increased(P < 0.05).(6)In terms of Protein expressions of NLRP3,GSDMD,Pro-Caspase-1,Pro-IL-1β,Caspase-1and ASC in synovium of rats in each group: Compared with sham operation group,the expression levels of NLRP3,GSDMD,Pro-Caspase-1,Pro-IL-1 β,Caspase-1 and ASC proteins in synovial membrane of model group were significantly increased(P < 0.05).Compared with model group,the expression levels of NLRP3,GSDMD,Pro-Caspase-1,Pro-IL-1β,Caspase-1 and ASC proteins in synovium of treatment group and control group were significantly decreased(P < 0.05).Compared with the treatment group,the expression levels of caspase-1,ASC,GSDMD and Pro-IL-1β in the control group were increased(P <0.05).While the expression levels of NLRP3 and Pro-Caspase-1 showed no statistical difference(P > 0.05).Conclusion:1.Document Research: In the treatment of different TCM syndromes of knee osteoarthritis,Electroacupuncture therapy and Electroacupuncture point-through therapy are mainly local acupoint selection.The largest number of acupoints is the bladder meridian,the largest frequency of acupoints are Stomach meridian of foot-yangming.For the treatment of different TCM syndromes,Electroacupuncture therapy and Electroacupuncture point-through therapy are relatively standardized and unified.In order of frequency,Dubi,Neixiyan,Xuehai,Yanglingquan,Liangqiu,Zusanli,Yinlingquan,Ashi and Heding appeared more frequently.Combined with the characteristics of anatomical position of knee and the method of Acupuncture therapy,Dubi through Neixiyan and Xuehai through Liangqiu were selected as the treatment methods.Through the literature study of Electroacupuncture treatment,theoretical support is provided for the clinical treatment of Knee-osteoarthritis,and the clinical efficacy of Electroacupuncture treatment for Knee-osteoarthritis needs to be further verified by clinical trials.2.Clinical Research: The clinical effect of Electroacupuncture point-through therapy on Knee-osteoarthritis is safe and reliable.It can effectively improve the symptoms of pain,swelling and stiffness of patients,improve their daily life ability,social life ability and participation in social entertainment,and improve their life and quality of life,which is worthy of further clinical application.In the treatment of Knee-osteoarthritis,Electroacupuncture point-through therapy can improve knee effusion,synovium thickness and synovium blood flow.The mechanism of its effect on synovium needs to be further explored in basic experiments.3.Fundamental research: Electroacupuncture point-through therapy can effectively inhibit NLRP3/caspase-1/IL-1β signaling pathway,thus reducing secretion of inflammatory cytokines IL-1β and IL-18,and synovial inflammation.To a certain extent,it can inhibit the occurrence of inflammatory cascade and pyroptosis of synovial cells.This indicates that the above treatment methods can effectively alleviate synovitis and have significant effects on delaying the progression of Knee-osteoarthritis. |
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