| Purpose:Research Objectives: To explore the effects of Yougui Pill on Rap1 signaling pathway,observe the expression of knee cartilage cells and inflammatory factors in serum of rats by intervening Yougui Pill in model rats with KOA knee osteoarthritis,and explore the expression of Yougui Pill on this pathway and the mechanism of action in preventing and treating knee osteoarthritis.To provide the theoretical basis for treatment.Research Methods:1 Establishment and grouping of KOA rat model: After 5 days of adaptive feeding,72 SPF SD rats(male and female)were randomly divided into 6 groups with 12 rats in each group by random number method.The left knee joint of rats was modulated by modified Hulth method.Anti-infection treatment was carried out for 3 days after operation,and wound healing was driven for 7 weeks.2 Model identification: 7 weeks after expulsion,a normal rat was randomly selected from the sham operation group and the model group to observe the morphology of cartilage and evaluate whether the modeling was successful.3 Drug intervention: sham operation group and model group were given distilled water intragastric administration.The other groups used body surface area method to calculate the daily dose required.4 Sampling: After anesthesia,blood was taken from abdominal aorta and centrifuged,and the separated serum and plasma were stored for future use.The cartilage dissection of knee joint was placed in PE tube and stored in-80℃ ultra-low temperature refrigerator for future use.5 Morphological examination of rat chondrocytes: HE staining was used to observe the pathological changes of rat chondrocytes.6 Observation and detection of chondrocyte indicators: wsetern-blot analysis of RAP1,RAF,MEK1/2,ERk1/2 and CREB protein expression in cartilage tissues of rats in each group,and analysis and comparison of the data of each group.The results:1 Effects of Yougui Pill on the contents of IL-1β,TNFα and IL-18 in serum of KOA rats:The contents of IL-1β,TNFα and IL-18 in serum of KOA rats in model group were significantly higher than those in sham operation group,which proved that the modeling was successful.Compared with model group,the expressions of IL-1β,TNFα and IL-18 in serum of Yougui Pill high,medium and low dose group and Western medicine group were significantly decreased,and the differences were statistically significant.2 Effects of Yougui Pill on the protein expressions of RAP1,RAF,MEK1/2 and ERK1/2 in articular cartilage of KOA rats: The protein expressions of RAP1,RAF,MEk1/2 and ERk1/2in articular cartilage of rats in model group were significantly increased compared with sham operation group;The protein expression of Yougui Pill high-dose group,Yougui Pill medium-dose group,Yougui Pill low-dose group and Western medicine group was lower than that of model histone.3 Effects of Yougui Pill on the expression of CREB,MMP3,MMP13 and ADAMTS proteins in articular cartilage of KOA rats: The expression of CREB,MMP3,MMP13 and ADAMTS proteins in articular cartilage of rats in model group was significantly increased compared with that in sham group.The protein expression of Yougui Pill high-dose group,Yougui Pill medium-dose group,Yougui Pill low-dose group and Western medicine group was lower than that of model histone.4 Effects of Yougui Pill on the expression of C-FOS,C-Myc and C-Jun proteins in articular cartilage of KOA rats: The expression of C-FOS,C-Myc and C-Jun proteins in the model group was significantly increased compared with that in the sham operation group;The protein expression of Yougui Pill high-dose group,Yougui Pill medium-dose group,Yougui Pill low-dose group and Western medicine group was lower than that of model histone.Conclusion:1 Yougui Pill can reduce the contents of IL-1β,TNFα and IL-18 in serum of rats and improve inflammation.2 The mechanism of Action of Yougui Pill in treating KOA is related to the expression of RAP1/RAF/MEK/ERK/CREB signaling pathway. |
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