Potential Biomarkers Of KOA Damp-heat Syndrome And Intervention With Simiaosa

Objective: Explores the potential biomarkers of KOA damp-heat syndrome and preliminarily studies the intervention effect of Simiao Powder.Methods:(1)The literatures on animal models of damp-heat syndrome from January 2000 to February 2022 were searched in China knowledge Network,Wanfang Medical Network,VIP Journal and Pub Med,and text information mining was carried out from the aspects of animal spec ies,intervention factors,related diseases,evaluation indicators and so on.(2)The rats were randomly divided into five groups: group A(sham operation),group B(KOA model),group C(KOA+ damp-heat),group D(KOA+ damp-heat + Simiao powder)and group E(KOA+ damp-heat + aconite decoction).Combined with the results of literature research on damp-heat syndrome model,groups C,D and E used Sodium iodoacetate combined with damp-heat environment to prepare KOA damp-heat syndrome combined animal model.The r at model of damp-heat syndrome of KOA was established by Simiao Powder and Wutou decoction respectively.The animal model of KOA damp-heat syndrome combined with disease and syndrome was verified from the general condition of rats,pathological examination and analysis and Mankin,s score.(3)The rats were randomly divided into A(sham operation group),B(KOA model group),C(KOA+ damp-heat group)and D(KOA+ damp-heat + Simiao powder group).According to the evaluation index of damp-heat syndrome model pro vided by literature research,the contents of TNF-α,IL-1β,IL-17 A,HSP-70,HIF-1α in serum,NA+-K+-ATP enzyme,Ca2 +-Mg2 +-ATP enzyme,succinate dehydrogenase and lactate dehydrogenase in liver were detected by ELISA,and the expression of HSP-70 and HIF-1 α in knee cartilage tissue was detected by immunohistochemical method.To explore the potential biomarkers of KOA damp-heat syndrome and to explore the intervention effect of Simiao Powder.Results:(1)The animal species of damp-heat syndrome animal model is mainly SD rats.The most common intervention factor of damp-heat syndrome is damp-heat environment,the highest frequency of temperature is 32 ℃,and the highest frequency of humidity is 90%.The study of animal model of damp-heat syndrome is mainly based on the model of combination of disease and syndrome.The study of syndrome model is mainly spleen-stomach damp-heat syndrome and febrile disease damp-heat syndrome,while the disease and syndrome com bined with animal model are mostly digestive system diseases and metabolic diseases.At present,the animal model of damp-heat syndrome is evaluated from three parts: symptom,physical sign and laboratory index.Symptom is the most important evaluation met hod.Damp-heat syndrome model is closely related to inflammatory index,energy metabolism index,HSP-70,HIF-1 α and so on.(2)C(KOA+ damp-heat group),D(KOA+ damp-heat + Simiao powder group),E(KOA+ damp-heat + aconite decoction group)rats showed diff erent degrees of dispirited spirit,happy pile,reduced food intake,reduced drinking water,sparse hair,sticky and greasy stool,increased defecation times,yellow urine and other damp-heat syndrome symptoms compared with A(sham operation group)and B(KOA model group)after damp-heat stimulation.After intragastric administration of Simiao Powder,the symptom of dampness-heat syndrome in group D was significantly better than that in group C,but not in group E.In Mankin’s,the score of group A was 0-1,that of group B and D was 4-7,which was the early and middle stage of KOA,that of group C and E was 7-11,which belonged to the middle and late stage of KOA.The Mankin’s score of group B,C,D and E was significantly higher thanthat of group A,the Man kin’s score of group C and E was significantly higher than that of group B,and the Mankin’s score of group D was significantly lower than that of group C and E.(3)Compared with A(sham operation group),the contents of NA+-K+-ATP enzyme,Ca2 +-Mg2 +-ATP enzyme,succinate dehydrogenase and lactate dehydrogenase in liver in group B(KOA model group),C(KOA+ damp-heat group)and D(KOA+ damp-heat + Simiao powder group)were significantly higher than those in group B(P < 0.0l).After treatment with Simiao powder,the index of energy metabolism in group D was significantly lower than that in group C(P < 0.05).The contents of TNF-α,IL-1β,IL-17 A,HSP-70 and HIF-1α in serum of rats in group B,C and D were significantly higher than those in group A,while the contents of TNF-α,IL-1β,IL-17 A,HSP-70 and HIF-1α in group C were significantly higher than those in group B and D.There were positive expressions of HIF-1α and HSP-70 in knee joints of all groups,a little expression of HIF-1α and HSP-70 in group A,pa rtial expression of HIF-1α and HSP-70 in group B,a large amount of expression of HIF-1α and HSP-70 in group C,and the expression of HIF-1α and HSP-70 in group D was significantly lower than that in group C.Conclusion:(1)The animal model of damp-heat syndrome has many intervention factors and a variety of modeling methods,each of which has its own advantages and disadvantages.Constantly refining the experimental variables and establishing an objective and reasonable evaluation system of damp-heat syndrome animal model is the key link in the research of damp-heat syndrome animal model.(2)Using knee joint injection of sodium iodoacetate combined with damp-heat external environment stimulation,the progress of KOA was faster and more serious than that of simple KOA model.The expression of KOA damp-heat syndrome combined with human KOA damp-heat obstruction syndrome was similar to that of human KOA damp-heat obstruction syndrome.based on the method of testing syndrome,the rat model of KOA damp-heat syndrome combined with damp-heat syndrome was established by selecting Simiao powder and Wutou decoction.It was found that Simiao Powder could significantly improve the Mankin’s score of rats with KOA damp-heat syndrome and inhibit histopathological changes a nd the degree of cartilage destruction.(3)KOA damp-heat syndrome is related to the upregulation of inflammatory cytokines TNF-α,IL-1β,IL-17 A and tissue HIF-1α and HSP-70,and the increase of liver energy metabolism indexes such as NA+-K+-ATP enzyme,Ca2 +-Mg2 +-ATP enzyme,succinate dehydrogenase and lactate dehydrogenase,which play an important role in judging KOA damp-heat syndrome.KOA damp-heat syndrome model is a potential marker.The effect of Simiao Powder on the rat model of KOA damp-heat syndrome may be through down-regulating the contents of TNF-α,IL-1β,IL-17 A,HIF-1α and HSP-70 in serum and reducing the index of energy metabolism.