Effect of Calcitonin-Mediated Expression of CD36 and IL-17 on Traumatic Osteoarthritis
Objective To investigate the protective effect of calcitonin on interleukin-1β(IL-1β)-induced chondrocytes,and further to study its potential mechanisms.Methods From July 21 to November 10,2022,human chondrocytes were treated with different concentrations of calcitonin,and the optimal concentration of calcitonin was selected by cell counting kit CCK 8(CCK-8)assay for follow-up experiments.Human chondrocytes were divided into control group,disease group and treatment group.The cell activity was detected by CCK-8 assay.The expression of CD36 and the production of intracellular reactive oxygen species(ROS)were analyzed by flow cytometry.The levels of superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by enzyme labeling method.The mRNA relative expression levels of calcitonin,interleukin-17(IL-17),matrix metalloproteinase 13(MMP13)and typeⅡ collagen(Col Ⅱ)were detected by quantitative real-time polymerase chain reaction(qRT-PCR).Western blot was used to detect the expression levels of NOD-like receptor heat protein domain associated protein 3(NLRP3),gasdermin-D(GSDMD)and GSDMD-N.Results It was found that the activity of human chondrocytes was concentration-dependent on calcitonin.Therefore,50 nM concentration of calcitonin was selected for subsequent experiments.In the disease group,the relative expression of Col Ⅱ mRNA was(0.47±0.06),the positive rate of CD36 was(0.17±0.02)%,and SOD was(151.14±12.26)U/mL,lower than control group(1.00±0.09),(1.50±0.16)%,(242.33±21.17)U/mL(P<0.05).Compared with the control group,the relative mRNA expression of MMP13 and IL-17,mean fluorescence intensity(MFI)of ROS and MDA content of the disease group were increased.Meanwhile,the expression levels of NLRP3 and GSDMD were increased,while the expression levels of GSDMD-N were decreased.However,the administration of calcitonin reversed the IL-1β-induced changes in chondrocytes.Pearson analysis showed that calcitonin was positively correlated with CD36 expression(r=0.922,P=0.001),negatively correlated with IL-17 expression(r=-0.881,P=0.002),and negatively correlated with CD36 expression level of IL-17(r=-0.650,P=0.023).Conclusion All those results suggested that calcitonin could protect IL-1β-induced chondrocytes damage,and its mechanism is related to mediating the expression of CD36 and IL-17,and alleviating the stress and inflammation of chondrocytes.
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