泽兰合剂对膝骨关节炎模型膝关节损伤后滑膜炎症反应及凝血功能的作用机制研究OA

Objective To explore the molecular mechanism of Zelan Mixture in intervening in the stress response in knee osteoarthritis(KOA)model rats from the perspectives of inflammatory response and coagulation function.Methods Fifty-five SD rats were randomly divided into the control group,the model group,and the low-,medium-,and high-dose Zelan Mixture groups.In all groups except for the control group,KOA animal model was built by anterior cruciate ligament transection method.After successful modeling,the control group and the model group were given equivalent volume of normal saline by gavage,while the low-,medium-,and high-dose Zelan Mixture groups were given 1 mL,2 mL,and 4 mL Zelan Mixture by gavage,respectively.The administration was performed twice a day for 7 days.The abdominal aortic blood and knee synovium of SD rats in each group were collected on the first,third and seventh day after gavage intervention for relevant tests.HE staining was used to observe the condition of synovial tissue in SD rats;Synovial tissue morphology score were used to evaluate the morphological change of synovial tissue;RT-qPCR was used to detect the relative expression of inflammatory-related genes IL-6,TNF-α,and NF-κB p65 in synovial tissue of SD rats in each group;Western blot was used to detect the relative expression of inflammatory-related proteins IL-6,TNF-α,and NF-κB p65 in synovial tissue of SD rats in each group;ELISA was used to detect the content of IL-6,TNF-α,ICAM,VCAM,and TXA2 in the serum of SD rats in each group.Results ①Synovial tissue morphology score:Compared with the model group,the synovial tissue morphology score of the high-dose Zelan Mixture group was significantly lower(P<0.05);②RT-qPCR:Compared with the model group,the relative expression levels of IL-6,TNF-α,and NF-κB p65 mRNA in synovial tissue of the medium-and high-dose Zelan Mixture groups were significantly lower(P<0.01);③Western blot:Compared with the model group,the relative expression levels of IL-6,TNF-α,and NF-κB p65 protein in synovial tissue of the medium-and high-dose Zelan Mixture groups were significantly lower(P<0.05);④ELISA:Compared with the model group,the serum contents of IL-6,TNF-α,ICAM,VCAM,and TXA2 in the low-,medium-,and high-dose Zelan Mixture groups were significantly lower at seventh day after surgery(P<0.05).Conclusion Zelan Mixture can alleviate the inflammatory indicators and improve the hypercoagulable state in KOA model rats,regulate the related expression of inflammatory-related genes and proteins in synovial tissue,and regulate the levels of inflammatory factors and coagulation factors in serum,thereby ameliorating the stress response in KOA model rats.The mechanism of action may be related to the reduction of inflammatory response and improvement of hypercoagulable state.