| Objective:In order to sort out the application of the classic prescription Duhuo Jisheng Decoction in modern medicine,and explore the current research status,development trend and hotspots of Duhuo Jisheng Decoction in the treatment of Knee Osteoarthritis(KOA),and to evaluate the advantages and disadvantages of various treatment methods.And to explore the effect of Jiawei Duhuo Jisheng Mixture(JDJM)on the Wnt/β-catenin signaling pathway of KOA synovium and its protective effect on KOA joint synovium and cartilage.Methods:1.Analysis of Modern Medication Characteristics of Duhuo Jisheng Decoction Based on Visualization Tools and Research Hotspots on Prevention and Treatment of KOA.(1)Retrieve 5 databases to obtain literature,use descriptive statistical analysis method to summarize relevant literature data on the modern application of Duhuo Jisheng Decoction,and describe and analyze the year of publication,journal,application dosage form,and treatment of diseases of the literature.(2)Using the method of co-occurrence analysis,the author,organization,and keywords of the literature are analyzed to find out the important researchers,core teams,and research institutions in this field.(3)The method of cluster analysis is applied to cluster the keywords in the relevant literature of Duhuo Jisheng Decoction in the treatment of KOA,find out the basic knowledge structure in this research field,and draw a timeline map to intuitively display the evolution process of this research field.(4)By using the method of keyword emergence analysis,the keywords in the relevant literature of Duhuo Jisheng Decoction in the treatment of KOA were analyzed,and the current research status,development trend and hotspots in this research field were explored.2.Clinical evaluation of Duhuo Jisheng Decoction in the treatment of KOA based on Network Meta-analysis.On the basis of part 1 of the literature search,we should screen and read the full text to obtain the information needed to build the data set.After the literature quality evaluation was included,the total effectiveness of different interventions for KOA was evaluated by using the network relationship diagram,and then the network evidence relationship diagram and the sorting probability bar chart of each outcome index were drawn by using R language to evaluate various interventions.3.JDJM regulates the Wnt/β-catenin signal pathway of rabbit KOA synovial fibroblasts in vitro.The rabbit model with KOA was established by using the improved Hulth modeling method,and the synovial fibroblasts of rabbits with KOA were isolated and cultured under sterile conditions after identification.With different volume fractions of JDJM,the expression levels of MMP-7 and Cyclin D1 in synovial fibroblasts culture supernatant were detected by ELISA,the expression of Wnt3 a,β-catenin,and COX-2 protein in synovial fibroblasts was detected by Western Blot,and the expression of Wnt3 a,β-catenin,MMP-7,Cyclin D1 and COX-2 m RNA in synovial fibroblasts were detected by RT-PCR,to explore the effect of modified JDJM on the signal pathway of Wnt/β-catenin in KOA synovial fibroblasts.4.JDJM regulating rabbit KOA synovial Wnt/β-catenin signal pathway in vivo.Sixty New Zealand healthy white rabbits were randomly divided into blank control group(15 rabbits)and model control group(45 rabbits).After model establishment,the model control group was randomly divided into model control group(15 rabbits),positive control group(15 rabbits)and traditional Chinese medicine group(15 rabbits).They were respectively given physiological saline,celecoxib and JDJM for intervention,and were killed 2,4 and 8 weeks after gavage.Under the light microscope,the microstructure of knee joint synovium was observed.Western blot method was used to detect the protein expression of joint synovium Wnt3 a,β-catenin,MMP-7,Cyclin D1 and COX-2,and RT-PCR method was used to detect the m RNA expression of joint synovium Wnt3 a,β-catenin,MMP-7,Cyclin D1 and COX-2,to explore the protective effect of JDJM on rabbit KOA cartilage and synovium and its effect on synovium Wnt/β-catenin signal pathway.Results:1.Analysis of Modern Medication Characteristics of Duhuo Jisheng Decoction Based on Visualization Tools and Research Hotspots on Prevention and Treatment of KOA.A total of 1603 papers were included in this study.In the past ten years,the maximum number of papers published in 2017 was 202;The relevant documents included were published in 260 journals,of which 66 were core journals,and the maximum number of articles published in the journal Clinical Research of Traditional Chinese Medicine was 85.The most commonly used drug form of Duhuo Jisheng Decoction is decoction;The treatment of diseases with Duhuo Jisheng Decoction involves orthopedics,internal medicine,gynaecology,ophthalmology,dermatology,etc.,of which 475 articles have been applied to KOA at most.Through further analysis of 475 documents,it is found that many research teams have been formed in this field,among which the top three influential teams are those centered around Professor Liu Xianxiang,Professor Lu Min and Professor Chen Shaoqing.Among the research institutions,Fujian University of Traditional Chinese Medicine Research Institute of Integrated Traditional and Western Medicine,Fujian University of Traditional Chinese Medicine and Fujian Provincial Key Laboratory of Integrated Traditional and Western Medicine for Senile Diseases ranked the top three.The First Affiliated Hospital of Hunan University of Traditional Chinese Medicine first published relevant documents,and the links between various scientific research teams were not close.The key words co-occurrence analysis shows that the research type in this field is mainly clinical observational research;Pain score and inflammatory factor level are the most commonly used indicators for efficacy evaluation;Acupuncture and moxibustion,joint cavity injection or NSAIDs are the most commonly studied combination therapies;Chondrocytes are the most frequently studied objects in experiments.Eight key word clusters were obtained from the study,namely Cluster 0(chondrocyte),Cluster 1(pain score),Cluster 2(celecoxib),Cluster 3(meloxicam),Cluster 4(acupuncture and moxibustion),Cluster 5(CRP),Cluster 6(clinical experience),Cluster 7(sodium hyaluronate);A total of 14 emergent keywords were obtained from the keyword emergent analysis,including clinical observation(2012-2013),clinical experience(2012-2013),traditional Chinese medicine fumigation(2012-2016),traditional Chinese medicine external application(2012-2013),nimesulide(2012-2013),traction(2012-2014),sodium hyaluronate(2013-2016),functional exercise(2013-2016),synovitis(2014-2016),exercise therapy(2015-2017),arthralgia syndrome(2016-2018)Inflammatory factors(2019~2021),mechanism of action(2019~2021),bone metabolism(2019~2021).2.Clinical evaluation of Duhuo Jisheng Decoction in the treatment of KOA based on Networkmeta-analysis.A total of 37 literatures were included in this study,including 1 English literatures and 36 Chinese literatures.37 literatures reported the effective rate of treatment,and 15 literatures reported adverse reactions.There are 17 interventions in 37 literatures,They are “Duhuo Jisheng Decoction ”,“glucosamine”,“celecoxib capsule”,“Duhuo Jisheng Decoction +acupuncture and moxibustion”,“diclofenac sodium”,“ibuprofen”,“ Duhuo Jisheng Decoction +glucosamine”,“meloxicam”,“sodium hyaluronate”,“ Duhuo Jisheng Decoction +functional exercise”,“ Duhuo Jisheng Decoction +sodium hyaluronate”,“ Duhuo Jisheng Decoction +external application/fumigation”,“diaceretin”,“ Extracorporeal shockwave therapy ”,“indomethacin”,“acupuncture and moxibustion”,“ Duhuo Jisheng Decoction +shock wave”;By comprehensively comparing the PASI score and the OR value of the total effective rate of different interventions,The advantages and disadvantages of the efficacy and safety are ranked as follows: D(Duhuo Jisheng Decoction +acupuncture and moxibustion)>J(Duhuo Jisheng Decoction +functional exercise)>L(Duhuo Jisheng Decoction +external application/fumigation)>K(Duhuo Jisheng Decoction +sodium hyaluronate)>Q(Duhuo Jisheng Decoction+ ESWT)>G(Duhuo Jisheng Decoction+glucosamine)>N(ESWT)>A(Duhuo Jisheng Decoction)>P(acupuncture and moxibustion)>I(sodium hyaluronate)>M(double acerhine)>O(indomethacin)>B(glucosamine)>F(ibuprofen)>E(diclofenac sodium)>C(celecoxib capsule)>H(meloxicam).3.JDJM regulates the Wnt/β-catenin signal pathway of rabbit KOA synovial fibroblasts in vitro.(1)The KOA model of rabbit can be successfully reproduced 5 weeks after operation.(2)ELISA test results showed that the content of Cyclin D1 and MMP-7 protein in different volume fractions of JDJM serum group was significantly lower than that in the same volume fraction of normal serum group after 48 and 72 hours of incubation;After 72 hours of incubation,the content of cyclin D1 and MMP-7 protein in the JDJM serum group with 20% volume fraction was the lowest(P<0.05),and there was no significant difference compared with the inhibitor group(P>0.05).(3)Western blot test results showed that after 48 and 72 hours of incubation,the content of Wnt3 a,β-catenin and COX-2 protein in different volume fractions of JDJM serum group was significantly lower than that in the normal serum group with the same volume fraction(P<0.05).There was a difference between the content of β-catenin and COX-2 protein in different volume fractions of JDJM serum group(P<0.05).The content of JDJM serum group with 20% volume fraction was less than that of JDJM serum group with 10% volume fraction was less than that of JDJM serum group with 5% volume fraction.(4)RT-PCR results showed that the expression levels of Wnt3 a,β-catenin,Cyclin D1,MMP-7 and COX-2 m RNA in different volume fractions of JDJM serum group were significantly lower than those in the same volume fraction of normal serum group after 48 and 72 hours of incubation(P<0.05),and there were significant differences between the β-catenin and COX-2 m RNA groups in different volume fractions of JDJM serum group(P<0.05),of which the m RNA expression level in the 20% volume fraction of JDJM serum group was significantly lower than that in the other volume fractions of JDJM serum group(P<0.05).4.JDJM regulating rabbit KOA synovial Wnt/β-catenin signal pathway in vivo.(1)After successful modeling,the rabbit’s mental state became worse,the amount of food consumption and activity decreased,and the thigh of the modeling side curled up protectively.From 2 days to 5 weeks after modeling,the mental state gradually improved,the amount of food consumption and activity gradually approached normal,and some still had claudication.(2)The diameter of rabbit knee joint increased after successful modeling(P<0.05);After JDJM,it gradually decreased and approached the normal level after 4 weeks(P>0.05).(3)After successful modeling,synovial hyperplasia of knee joint was observed by gross observation,with bright yellow and white color and hard texture,and a large amount of blood stasis was observed;With time,the color of synovial membrane is dark yellow,and fibrous hyperplasia can be seen,and the texture hardness increases.After intragastric administration of JDJM,the color of synovial membrane became lighter and the proliferation improved.The surface of the condylar cartilage in the articular cartilage is extremely rough,the touch is not smooth,the color of the cartilage is dark and opaque,the cartilage is thinned or missing locally,the bottom is congested,and the joint fluid is turbid;After the intervention of traditional Chinese medicine,the surface is still flat,the touch smoothness is fair,and the cartilage is transparent.(4)After successful modeling,under the synovial tissue microscope,the distribution of surface cells was uneven,and the cells in the synovial lining layer proliferated in large numbers,up to 5-7 layers.The cell arrangement was obviously disordered,and a large number of inflammatory cells infiltrated,and blood vessels and fibrous tissues proliferated significantly.With the increase of time,the number of inflammatory cells increased,and blood vessels and fibrous tissues proliferated seriously.After the intervention of traditional Chinese medicine,the number of lining cells decreased,inflammatory cells decreased,and the proliferation of fibrous tissue gradually improved.Under the cartilage microscope,the thickness of cartilage becomes thinner,the cracks on the surface of cartilage are obvious,and some of them can reach the radiation area.The distribution of chondrocytes is irregular,the number of chondrocytes decreases,the tidal line disappears,and some of them can see blood vessels pass through.As time goes on,partial cartilage exfoliation and subchondral bone exposure can also be seen.After the intervention of traditional Chinese medicine,the surface of cartilage was rough and uneven,the proliferation of chondrocytes crept to the injured area,the thickness of cartilage became thinner,and the tidal line was blurred.With the increase of gastric perfusion time,cartilage repair is obvious,and the tide line is basically visible,but the overall thickness is still relatively thin.(5)After successful modeling,compared with the blank control group,the expression levels of Wnt3 a,β-catenin,Cyclin D1,MMP-7 and COX-2 m RNA in synovial tissue of each group were higher(P<0.05),and most of them tended to increase with time;After intragastric administration of JDJM,it gradually decreased,with statistical difference compared with the model control group(P<0.05),and no statistical difference compared with the positive control group and the blank control group(P>0.05).(6)After successful modeling,compared with the blank control group,the content of Wnt3 a,β-catenin,Cyclin D1,MMP-7 and COX-2 protein in synovial tissue of each group was higher(P<0.05).After intragastric administration of JDJM,the protein content of Wnt3 a,β-catenin,Cyclin D1,MMP-7 and COX-2 in the traditional Chinese medicine group was close to that in the blank control group after 2 weeks(P>0.05).Conclusion:1.Duhuo Jisheng Decoction still shows considerable advantages in the treatment of modern diseases,and its value should be fully explored to make contributions to the modernization of traditional Chinese medicine and the inheritance,innovation and development of traditional Chinese medicine.The number of documents published on Duhuo Jisheng Decoction has declined slightly after increasing year by year,and is now in a stable state,which means that the research in this field has gradually entered a stable development stage.2.Duhuo Jisheng Decoction is the most widely used in orthopedics,especially in the treatment of KOA.The research of Professor Liu Xianxiang of Fujian University of Traditional Chinese Medicine and Professor Lu Min of Hunan University of Traditional Chinese Medicine on the treatment of KOA with Duhuo Jisheng Decoction is relatively mature,which can be the first consideration for academic exchange and cooperation at home and abroad in this field.3.In the exploration of the mechanism of Duhuo Jisheng Decoction in treating KOA,the previous hot topic was clinical observation to confirm the clinical efficacy of this formula;The current hot topic is to verify the mechanism of action of this formula,mainly focusing on evaluating changes in cartilage;The next trend may shift towards the impact of synovial and “synovial – cartilage” interaction mechanisms on KOA,with experimental verification as the main research method.4.Duhuo Jisheng Decoction can effectively treat KOA with less side effects;In comprehensive therapy,Duhuo Jisheng Decoction combined with acupuncture and moxibustion has the best effect,while in single therapy,Duhuo Jisheng Decoction has significant therapeutic effect;Compared with single therapy,combined therapy can improve clinical efficacy.5.The KOA model of rabbit can be successfully established by the improved Hulth method for the study of relevant mechanisms.6.Wnt/β-catenin signal pathway is activated in KOA.And down-regulation of Wnt/β-catenin signal pathway can inhibit the proliferation and inflammatory reaction of synovial fibroblasts.7.Articular cartilage injury is related to synovium in KOA.8.JDJM can regulate Wnt/β-catenin signal pathway,improve KOA of rabbit symptoms,repair articular cartilage,inhibit synovial hyperplasia,and inhibit the secretion of inflammatory factors. |
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