Exploring The Mechanism Of Bushen Huoxue Prescription In Treating Knee Osteoarthritis Based On The Regulation Of Chondrocyte Apoptosis By NF-κB Signaling Pathwa

Objective:In this study,the effects of Kidney Booster on apoptosis and NF-κB pathway-related genes and protein expression were observed by intervening in knee osteoarthritis model mice,and to explore the possible targeting mechanism of Kidney Nourishing Blood Formula on delaying the effect of knee osteoarthritis.Methods:Twenty-four 6-week-old male mice with BALB/c were randomly divided into three groups:sham surgery group,KOA model group and traditional Chinese medicine group to construct a model of knee osteoarthritis by Hulth method by using the modified artificial induction model.After 1 week after surgery,gastric gavage was performed,and the drug group was gavaged with 0.243 g·kg-1·d-1 to replenish kidney and blood circulation,the volume of gavage was 10mL·kg-1·d-1,and the blank group and model group were given equal volumes of normal saline,and gavage was continuously administered for 4weeks.After 6 weeks,the morphological changes of cartilage tissue were observed by H&E staining and saffred solid green staining,and the expression rate of IL-6,INOS and COX2 inflammatory cytokines was detected by ELISA.PCR detected Collagen II.,Aggrecan,ADAMTS-5 proteins in knee tissue,and Western blot measured Bcl-2,Cleaved-Caspase-9,Bax and p-IκBα/IκBα,p-P65/P65 protein content.Results:(1)Joint chondrocytes in the model group and the sham surgery group had different degrees of pathological degeneration,chondrocytes arrangement disorder and hypertrophy,while in the traditional Chinese medicine group,the thickness of hyaline cartilage(HC)decreased compared with the model group,and the phenomenon of chondrocytes disorder and hypertrophy improved.Compared with the sham surgery group,the proteoglycan of cartilage in the model group was significantly reduced,and the loss of proteoglycans was partially reduced after the treatment of Kidney Nourishing Blood Soup.(2)Compared with the sham group,the expression level of Collagen II and Aggrecan in the knee joint tissues of mice in the model group decreased significantly,and ADAMTS-5 increased significantly.Compared with the model group,the expression of Collagen Ⅱ and Aggrecan in the traditional Chinese medicine group increased significantly compared with the KO A group,and the expression of ADAMTS-5 in the traditional Chinese medicine group decreased significantly,and the difference was statistically significant.(3)The concentrations of INOS,IL-6 and COX2 in each group of mice in the semester were compared,and the concentrations of INOS,IL-6 and COX2 were significantly increased in the model group compared with the sham group.Compared with the model group,the expression of INOS,IL-6 and COX2 in the serum of mice in the traditional Chinese medicine group decreased significantly.()Compared with the sham surgery group,the expression levels of Cleaved-Caspase-9,Bax and p-IκBα/IκBα and p-P65/P65 proteins in the knee joint tissues of mice in the model group were significantly increased,and the expression of Bcl-2 was significantly reduced;Compared with the model group,the expression of Cleaved-Caspase-9,Bax and p-IκBα/IκBα and p-P65/P65 proteins decreased significantly,and the expression of Bcl-2 increased significantly.Conclusion:1.Kidney supplementation and blood activation can effectively improve the inflammatory response of KOA mouse model and reduce cartilage damage.2.The mechanism of action of the kidney tonic and blood revitalization formula in the treatment of KOA is related to the expression of relevant factors and proteins that regulate apoptosis.