Based On The JAK/STAT Signaling Pathway To Explore The Mechanism Of Xinzhigushengchang Pills On Chondrocyte Osteoarthritis Model

Purpose:To investigate the modeling concentration of IL-1?-induced C-28/I2 osteoarthritis model in human chondrocytes;to screen the optimal effective concentration of New Bone Hyperplasia Pill medicated serum on C-28/I2 osteoarthritis model cells;to explore the effect of New Bone Hyperplasia Pill medicated serum on the secretion of IL-6 and TNF-? levels and the expression of JAK2 protein expression level and phosphorylation level and the expression of negative feedback regulatory protein SOCS1 in IL-1?-induced C-28/I2 osteoarthritis model at the effective concentration.Thus,to observe the effect of New Bone Hyperplasia Pill medicated serum on JAK-STAT signaling pathway and explore the mechanism of New Bone Hyperplasia Pill on osteoarthritis.Material and method:Four different concentration gradient groups of IL-1? were set to intervene C-28/I2,and the proliferation activity of C-28/I2 was detected by CCK-8 assay 24 hours later to select the optimal concentration for preparing osteoarticular chondrocyte models and prepare osteoarthritis model chondrocytes.Forty female New Zealand white rabbits aged 6 months(500±50g)were divided into two groups and intragastrically administered with New Bone Hyperplasia Pill and the same amount of saline for 2 weeks,respectively,and blood samples were collected from the abdominal aorta to prepare medicated serum for New Bone Hyperplasia Pill With 0 concentration as the control group,6different concentrations of New Bone Hyperplasia Pill medicated serum were set to intervene C-28/I2,and then the cell viability of each concentration group of New Bone Hyperplasia Pill medicated serum was determined by CCK-8 method,and the proliferation curve was drawn to select the optimal concentration of C-28/I2 medicated serum intervention.The secretion of TNF-? and IL-6 in the culture supernatant of C-28/I2 cells in each group was measured by ELISA;the protein expression and phosphorylation level of JAK2,as well as the expression of SOCS1 protein in each group of cells were detected by Western blot.Results:1 When the concentration of IL-1? was equal to 10 ng/ml,the OD value of C-28/I2 cells was statistically different from that of the control group(p < 0.05).2 When the medicated serum concentration was 10%,the OD value of C-28/I2 cells was statistically different from that of the control group(p < 0.05).3 Compared with the Mod group,the contents of inflammatory factors TNF-? and IL-6 were reduced and significantly different in the XZG group(p < 0.01).4 Compared with the Mod group,the expression of JAK2 and p-JAK2 relative protein in the XZG group was significantly different(p < 0.01);SOCS1 protein expression was increased and the difference was statistically significant(p< 0.05).Conclusion:1 IL-1? inhibited the proliferation of C-28/I2 cells,and the concentration of osteoarthritis model was 10 ng/ml.2 The serum containing New Bone Hyperplasia Pill can promote the proliferation of C-28/I2 cells,and the optimal concentration is 10%.3 New Bone Hyperplasia Pill can reduce the secretion of inflammatory factors IL-6 and TNF-? in C-28/I2 cells.4 New Bone Hyperplasia Pill can inhibit the expression of JAK2 protein and JAK2 phosphorylation and activate the expression of negative feedback protein SOCS1 thereby inhibiting the JAK/STAT signaling pathway.New Bone Hyperplasia Pill may inhibit the degeneration and apoptosis of chondrocytes by reducing the secretion of inflammatory factors IL-6 and TNF-?,inhibiting the expression of JAK2 protein and JAK2 phosphorylation and activating the expression of negative feedback protein SOCS1 to inhibit the JAK/STAT signaling pathway.