Clinical Efficacy Evaluation And Network Pharmacology Study Of Modified Guizhi Fuzi Decoction In The Treatment Of Knee Osteoarthritis Of Yang Deficiency And Cold Coagulation Type

Objective In this study,we designed a clinical controlled trial to observe and evaluate the clinical efficacy and safety of Modified Guizhi Fuzi Decoction in the treatment of Yang deficiency and cold condensation type of knee osteoarthritis,determined the regulatory mechanism of Modified Guizhi Fuzi Decoction in the body by Elisa assay,and applied the network pharmacology method to predict the potential mechanism of action of Modified Guizhi Fuzi Decoction,and initially supported the scientific of its clinical application.Methods1.Clinical observation research part Sixty-six invalids belonged to Yang deficiency and cold coagulation syndrome of knee osteoarthritis of in the outpatient programme and inpatient ward of Department of Rheumatology of Hohhot Mongolian Medicine Hospital were differentiated into two sets in parallel and randomly,30 patients in the treatment group and 30 patients in the control group.The treatment group was treated with Guizhi Fuzi Decoction combined with atractylodis rhizoma,mulberry parasitoids and eucommia ulmoides,1 dose/d.Glucosamine sulfate capsules was given to the matched group,at every turn 500 mg,thrice/d.Both groups were treated for 6 weeks.The visual analogue scale(VAS),Lequesne index score,WOMAC index score and Yang deficiency syndrome score were analyzed before and after treatment for 2,4 and 6weeks,and the changes of serum interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and matrix metalloproteinase-3(MMP-3)levels were observed in the two groups before and after treatment,and the clinical efficacy of patients in the two groups was also compared.2.Network pharmacology research part The active ingredients and targets of each drug in Modified Guizhi Fuzi Decoction were retrieved through TCMSP and BATMAN-TCM databases,and the relevant targets of Knee Osteoarthritis(KOA)were mined through Drug Bank,Gene Cards,OMIM,Dis Ge NET and TTD databases,and the common targets of drugs and disease were retrieved using Venn diagram online tool in Oebiotech Cloud Platform,we used STRING database and Cytoscape software to construct the protein interaction network(PPI)and perform topology analysis to obtain the core targets.Finally,GO and KEGG enrichment analyses were performed in the Metascape database with the common targets to derive the molecular functions,cellular components,biological processes and related signaling pathways that may be involved in the treatment of KOA with Modified Guizhi Fuzi Decoction,and to analyze the potential mechanism of Modified Guizhi Fuzi Decoction for the treatment of KOA.Results1.Clinical observation research part In the end,60 patients completed all the trial,30 each in the treatment group and the control group.Baseline data and VAS index,Lequesne index,WOMAC index,Yang deficiency symptom score,serum IL-1β,TNF-α and MMP-3 levels were compared between the two groups before medication,and the differences were not statistically significant(P > 0.05).The differences in VAS index,Lequesne index and WOMAC index scores were statistically significant(P < 0.05);the differences in Yang deficiency symptoms scores were statistically significant(P < 0.05);the differences in serum IL-1β,TNF-α and MMP-3 levels were statistically significant in both groups before and after medication(P < 0.05).The differences in VAS index,Lequesne index and WOMAC index between the two groups after treatment were not statistically significant(P > 0.05);the differences in Yang deficiency symptoms scores were statistically significant(P < 0.05);the differences in IL-1β,TNF-α and MMP-3 levels were not statistically significant(P > 0.05).The overall clinical effective rate of the treatment group was 86.7%,and the difference was not statistically significant compared with 83.3% in the control group(P > 0.05).No significant adverse effects were seen in either group,there was no statistically significant difference in blood routine,urine routine,liver function and kidney function in both groups before and after treatment(P > 0.05).2.Network pharmacology research part The search yielded 95 active ingredients of Modified Guizhi Fuzi Decoction,and the final obtained total of 57 active ingredients,corresponding to 271 targets after removing duplicates and non-target components.A total of 1440 relevant targets about KOA were obtained.A total of 121 drug-disease common targets were obtained by Venn diagram,corresponding to 44 major active ingredients.By constructing the PPI network diagram of Guizhi Fuzi Decoction in the treatment of KOA,it can be concluded that the treatment of KOA with Modified Guizhi Fuzi Decoction is related to 24 core targets.GO and KEGG enrichment analysis showed that there were 4659 GO ontology entries,,including 1752 for BP,2711 for MF,and 196 for CC.196 pathways were obtained from KEGG pathway enrichment analysis,and the treatment of KOA with Modified Guizhi Fuzi Decoction mainly involved PI3K-Akt,IL-17,TNF and other signaling pathways.Conclusion Modified Guizhi Fuzi Decoction can significantly relieve the symptoms of knee swelling,pain and stiffness,improve the function of joint movement,and reduce the score of Yang deficiency symptoms and the levels of IL-1β,TNF-α and MMP-3 in patients’ serum,which is a clear effect for the treatment of Yang deficiency and cold condensation type of knee osteoarthritis.The mechanism of action may be related to its inhibition of serum inflammatory factor release and secretion of matrix metalloproteinase.The route of action of component-target-pathway were complex and diverse of Modified Guizhi Fuzi Decoction in the treatment of KOA.The key active components such as quercetin,beta-sitosterol,kaempferol,beta-carotene,wogonin,Stigmasterol,18beta-Glycyrrhetinic Acid,Glycyrrhetinic Acid,18alpha-Glycyrrhetinic Acid,(+)-catechin may act IL6,ALB,TNF,AKT1,IL1 B,VEGFA,TP53,MMP9,PTGS2,CASP3 and other core targets synergistically through multiple pathways such as PI3K-Akt signaling pathway,IL-17 signaling pathway,TNF signaling pathway intervene and regulate the occurrence and development of KOA.This study provides theoretical support and clinical rationale for the clinical application of Modified Guizhi Fuzi Decoction in the treatment of KOA.