Clinical And Experimental Study Of Electroacupuncture In The Treatment Of Inflammation-mediated Osteoarthritis Cartilage Degeneration

Objective:1.Documentary research:To analyze the rules for acupoint selection of electroacupuncture in clinical treatment of knee osteoarthritis based on data mining technology to provide clinical guidance.2.Clinical experiments: A randomized controlled trial was used to verify the effects of electroacupuncture in the treatment of wind-cold-dampness knee osteoarthritis and effects on cell inflammatory factors.3.Basic research: To explore the potential mechanism of electroacupuncture in the treatment of inflammation-mediated osteoarthritis cartilage degeneration based on the P38 MAPK pathway.??Methods:1.Documentary research:The literatures in CNKI,Wanfang,VIP and Pub Med Data about acupuncture treatment of knee osteoarthritis from established until December 31,2018 were retrieved through computer and manual retrieval.The literatures database was established and the acupoints information including their channel tropism,locations specific acupoint attributes and syndrome acupoint frequency were analyzed by frequency and percentage.Based on the software of TCM inheritance auxiliary platform(V2.5),the common medication rules were searched out through analyzing the selected data by using Apriori algorithm in association analysis and the core prescription and combination of new prescription were obtained by using cluster analysis.2.Clinical experiments: 150 cases were randomly divided into treatment group and control group with 75 cases in each group.Treatment group was given electroacupuncture,a daily treatment,15 times for a period of treatment,the treatment course,rest two days after the end of the ground were two consecutive treatment period of treatment.Control group was given intraarticular injection of sodium hyaluronate,2ml each knee time,once a week for fiveweeks for a course.VAS score,WOMAC score,FTSST time were observed before and after treatment,so were the IL-1?,TNF-? contents by ELISA.The data obtained are used SPSS20.0 software analysis.3.Basic research:(1)Thirty healthy 2-month-old male SD rats were randomly divided into control group(normal group,24 rats)and experimental group(electro-acupuncture intervention,6 rats).Free drinking water,standard feed.The normal group did not give any intervention;The experimental group received electro-acupuncture intervention for 15 min/time.The remaining intervention conditions were the same as that of the experimental group 1.After 12 hours of fasting before blood collection and 3 hours of last electro-acupuncture treatment,the abdominal aorta blood was collected after anesthesia with 2% sodium pentobarbital(40mg/kg).The upper serum was collected after high-speed centrifugation and stored at-80 C for reserve at low temperature.(2)The knee osteoarthritic chondrocytes of healthy 4-week-old SD rats(10 rats)were obtained by mechanical-enzymatic digestion,and the optimal chondrocytes were screened and identified by differential adherence purification in vitro.The optimal time-effect of LPS-induced chondrocyte inflammation model was screened by ELISA after different concentrations and different time intervening.The expression of IL-1beta,IL-6 and TNF-alpha in each group after LPS-induced inflammation model was intervened by Qing(normal group,control group,model group,electro-acupuncture group for 15 minutes).The expression of P38MAPK?MEK3?MEKK3?c-myc?c-jun?c-fos s gene and the expression of P38MAPK?MEK3?MEKK3?c-myc?c-jun? c-fos protein in chondrocytes were detected by q PCR and Western-blot.The data obtained are used SPSS20.0 software analysis.Results:1.Documentary research:Total 385 points in 347 pieces of literature were included.Acupoints with high frequency included Waixiyan,Neixiyan,Xuehai,Yanglingquan,Liang-qiu,Zusanli,Yinlingquan,Ashixue,Heding and most of them located on lower extremity.The meridians for the treatment of KOA acupoints by electroacupuncture are mainly concentrated in the foot Yangming stomach and foot Taiyin spleen,and there are Ashi points and the knee and crane top in the outside Qi points;The specific points are more commonly selected,and the proportion of five points is higher;The points taken by different types of KOA are stilldominated by local points and meridians;Acupoints combination is mainly local acupuncture points.The core acupoints combination and new acupoints combination can provide reference for clinical medication,while their curative effects need to be further studied.2.Clinical experiments: Clinical data of two groups of cases comparable,it had no statistically significant difference(P>0.05).The treatment group,effective rate was 88.59%,the control group effective rate was 79.17%.The difference in total clinical efficiency between the two groups was statistically significant(P< 0.05);There was no significant difference in VAS score,total WOMAC score and individual score(pain,stiffness,degree of daily activity),contents of IL-1?,TNF-? between the two groups before treatment(P>0.05),they were comparable;Compared with pre-treatment,VAS score,total WOMAC score and individual score(pain,stiffness,degree of daily activity),contents of IL-1?,TNF-?,in both control and treatment groups were reduced obviously after treatment(P< 0.05).VAS score,total WOMAC score and individual score(pain,degree of daily activity)showed statistic difference in control and treatment groups after treatment(P< 0.05)while stiffness score of WOMAC and contents of IL-1?,TNF-? by ELISA showed no statistic difference(P>0.05).3.Basic research:(1)Chondrocyte in the best condition was selected as the second generation chondrocyte by differential adherence purification method;(2)ELISA showed that the expression of IL-1beta,IL-6 and TNF-alpha in the supernatant of chondrocyte was significantly higher in the model group than that in the blank serum group(p<0.05 or P<0.01),while the expression of IL-1beta,IL-6 and TNF-alpha in the control group,the experimental group were significantly lower than that in the model group(p<0.05 or P<0.01).(3)The results of q PCR and WB showed that the expression of P38MAPK? MEK3? MEKK3? c-myc?c-jun? c-fos in LPS-induced chondrocytes could be down-regulated after 15 minutes of EA intervention.Compared with the normal group,the levels of P38MAPK?MEK3?MEKK3?c-myc?c-jun?c-fos m RNA and protein in the model group were significantly higher(p<0.05 or P<0.01).Compared with the model group,the levels of protein and protein in the experimental group were significantly lower(p< 0.05 or P<0.01).Conclusion:1.Documentary research:The acupoints in electroacupuncture treatment of KOA is biased towards local points,meridians,and specific points.Acupoints with high frequencyincluded Waixiyan,Neixiyan,Xueha,Yanglingquan,Liangqiu,Zusanli,Yinlingquan,Ashixue,Heding,Mainly concentrated in the foot Yangming stomach and foot Taiyin spleen.The use of specific points is most prominent in the combination of five points.The points taken by different syndromes KOA are still dominated by local points and meridians.There are relatively few points for dialectical points,and they are also relatively inconsistent.This may be the place where acupuncture clinicians should think most.In fact,some distant points can highlight the characteristics of syndrome differentiation.For example,the prescription(Neixiyan,Waixiyan,Yinlingquan,Yanglingquan,Xueha,Zusanli,Sanyinjiao.Ashixue)takes into account the far part of the point at the same time as taking partial points,and can be used to relieve cold,dampness,and pain,suitable for cold wet type KOA,The clinical efficacy of this prescription will also be partially confirmed in clinical studies.The core acupoints combination and new acupoints combination can provide reference for clinical medication,while their curative effects need to be further studied.2.Clinical experiments: Clinical experiments: Electroacupuncture and intraarticular injection of sodium hyaluronate on wind-cold-dampness knee osteoarthritis has a clear therapeutic effect.There was no significant difference between the two groups in the relief of stiffness.The electroacupuncture group has obvious advantages in relieving pain symptoms,improving joint function and coordination and balance ability of the body.2.Both groups can effectively reduce the IL-1?,TNF-? of synovial fluid.Reducing inflammatory factors may be one of the mechanisms of electroacupuncture in the treatment of knee osteoarthritis.3.Basic research:(1)Serum after electro-acupuncture can reduce LPS-induced chondrocyte inflammation.Its mechanism is to down-regulate the expression of inflammation-related signal P38MAPK? MEK3? MEKK3? c-myc? c-jun? c-fos gene protein.It can play an anti-inflammatory role by inhibiting the expression of inflammatory factors such as IL-1beta,IL-6,TNF-alpha,thereby delaying the apoptosis of chondrocyte.