Experimental Study On The Effect Of Xitongkang On The TNF-MAPK-Fas Signaling Pathway And The Inhibition Of Apoptosis Of Chondrocytes In The Knee Joint Of Rats With KOA Mode

Objective:To explore the effects of "Xitongkang" on cartilage degeneration,cartilage tissue MAPK signaling pathway and serum related apoptosis proteins of knee osteoarthritis based on network pharmacology,and to explore the mechanism of "Xitongkang" on the apoptosis of cartilage cells of knee osteoarthritis,so as to provide a substantial laboratory basis and theoretical basis for clinical treatment and further research.Material and methods:1.Establishment and grouping of KOA rat model: Forty-eight SPF SD rats(half male and half female)were selected.After 7 days of adaptive feeding,the rats were divided into sham operation group,model group,"Xitongkang" group and celecxib group by random number table method.The right knee joint of rats was modeled after adaptive feeding.The sham operation group was sutured after opening the joint cavity of the right hind limb without any other operation.The other groups were modeled by modified Hulth method.The rats were forced to exercise for 8 weeks after surgery.The model was evaluated to determine the therapeutic intervention after the model was established.2.Model identification: 8 weeks after the drive,one normal rat was randomly selected from the sham operation group and the model group for observation,and the cartilage morphology was observed to evaluate whether the model was successfully constructed.3.Drug administration: The rats in sham operation group and model group were given normal saline by intragastric administration.Body surface area method was used to calculate the daily dose of drugs in the other experimental groups.4.Experimental materials: After anesthesia,blood was taken from abdominal aorta.After centrifugation,serum and plasma were separated and retained respectively To use;The knee cartilage was separated and placed in EP tubes and stored at-80℃ in ultra-low temperature refrigerator.5.Morphological detection of rat chondrocytes: toluidine blue and HE staining were used to observe the pathological changes of rat cartilage.Chondrocyte apoptosis was observed by TUNEL staining.Mankin score was calculated to evaluate cartilage degeneration.6.Observation and detection of chondrocyte indicators: The expressions of JNK1/2,P38,ERK1/2,p-JNK1/2,p-P38,p-ERK1/2 and the changes of related apoptotic proteins such as Fas and Jag1 in rat cartilage tissues in each group were detected by Western-blot analysis,and the data of each group were analyzed and compared.Results:1.Compared with the sham operation group,the score of the model group was significantly higher(P<0.01).Compared with model group,the scores of Xitongkang group and celecoxib group were significantly decreased(P<0.01).2.The effect of Xitongkang on the contents of IL-1β and TNFα in the serum of KOA rats: the contents of IL-1β and TNFα in the serum of KOA rats in the model group were significantly higher than those in the sham operation group,which proved that the model was successfully constructed.Compared with model group,the expressions of IL-1β and TNF-α in serum of rats in Xitongkang group and celecoxib group were significantly decreased,and the difference was statistically significant.3.Effect of Xitongkang on JNK1/2,P38,ERK1/2,p-JNK1/2,p-P38,p-ERK1/2 protein expression in articular cartilage of KOA rats: Compared with sham operation group,JNK1/2,P38,ERK1/2,p-JNK1/2,p-P38,p-ERK1/2 protein expressions were significantly increased in model group.The expression of histone in Xitongkang group and celecoxib group decreased compared with the model.4.The effect of Xitongkang on the expression of Fas and Jag1 protein in the articular cartilage of KOA rats: compared with the sham operation group,the expression of Fas and Jag1 protein in the cartilage of rats in the model group was significantly increased;Fas and Jag1 protein expressions in Xitongkang group and celecoxib group were decreased compared with model group.Conclusion:1.Xitongkang can reduce the contents of Fas and Jag1 in knee chondrocytes of rats,and reduce the apoptosis of chondrocytes.2.The mechanism of action of Xitongkang in treating KOA is related to its regulation of TNF-MAPK-Fas signaling pathway related factors.