Analysis Of The Components Of Xuanhusuo And Its Network Pharmacology Research On The Mechanism Of Action Of Osteoarthritis

Osteoarthritis(OA)refers to a chronic illness of the joints and bones featured by progressive articular surface damage by articular cartilage injury and the imbalance of cartilage matrix synthesis and degradation.Currently,most OA treatment strategies,including conservative treatment,drug therapy and surgical treatment,mainly focus on reducing symptoms and preventing injury of joints.Patients usually take NSAIDs and cartilage protection agents,however,reports show that long-term application will cause side effects on the gastrointestinal tract.In terms of the concept of traditional Chinese medicine(TCM),OA can be classified as producing "arthromyodynia"(blockage syndrome,Bi syndrome,or Bi Zheng),which is caused by qi and blood obstructed and tendon nutrient deficiency.Thus,TCM focuses on tonifying kidney,promoting blood circulation,clearing damp and relieving pain to treat OA.The use of Xuan Hu Suo San(XHSS)was recorded in Pu Ji Fang,which is prepared from four herbs including Corydalis yanhusuo W.T.Wang(Yan Hu Suo),Achyranthes bidentata B1.(Niu Xi),Angelica sinensis(Oliv.)Diels(Dang Gui),and Psoralea corylifolia L.(Bu Gu Zhi)to treat deficiency of liver and kidney,blood clotting and qi stagnation,lumbar and leg ache,represents the holism and compatibility of TCM on OA with the complementary relations among four herbs.XHSS has been applied in clinical widely and recorded in the Orthopedic Prescription Prescription Management System of Wangjing Hospital of China Academy of Chinese Medical Sciences,however,there have been no relevant researches about XHSS especially its mechanism on OA until now,which restrict its clinical application and is necessary to do further research.Thus,the following four aspects were studied:Firstly,effect of XHSS and its disassembled prescriptions on dissolution of psoralen and isopsoralenThe dissolution of psoralen and isopsoralen in XHSS and its disassembled prescriptions were determined by HPLC.Results showed that dissolution of psoralen and isopsoralen had great changes among XHSS and its disassembled prescriptions,Angelicae Sinensis and Achyranthis Bidentatae were helpful to the dissolution of psoralen and isopsoralen from Psoralea corylifolia,while Corydalis yanhusuo could inhibit their dissolution,which provides basis for further revealing the compatibility rules of XHSS.Secondly,ultra high performance liquid chromatography with tandem mass spectrometry for identification of main constituents in XHSSUltra high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry was applied to rapidly separate and identify the main chemical ingredients in XHSS for the first time.Finally,with the optimized separation and detection method,a total of 57 compounds were simultaneously separated within 13 min and confirmed by comparing retention time and MS data with reference standards,database or reference literatures,which provides basis for the study of pharmacokinetics.Thirdly,network pharmacology approach to uncover the pharmacological mechanism of XHSS on osteoarthritisNetwork pharmacology approach was chosen to study the pharmacological mechanism of XHSS on OA,and the pharmacology networks were established based on the relation amongst four herbs in XHSS,compounds targets,OA targets,and protein-protein interaction.Finally,the pathway enrichment analysis revealed the significant bioprocess networks of XHSS on OA were regulation of inflammatory,IL-1β production and NO biosynthetic process,response to cytokines or estrogen stimulus and anti-apoptosis.The comprehensive network pharmacology approach developed by our research had revealed the connection of four herbs in XHSS with corresponding compounds targets,and OA pathway systems for the first time,which provides basis for further research.Fourthly,effects of XHSS on the cartilage matrix of OA ratsTo investigate the effects of XHSS on the cartilage matrix of OA rats,male SD rats were randomized and treated by anterior cruciate ligament transection and medial meniscus resection to create OA models.The rats were sacrificed after treated with XHSS for three and six weeks,their knee joints were stained with HE and Masson to observe the cartilage injury and collagen degradation,respectively.The results showed that XHSS could obviously suppress the cartilage injury and collagen degradation(P<0.05),which initially proves the potential therapeutic efficiency of XHSS on OA and provides basis for further vitro experiments and verification experiments of network pharmacology prediction.