Study On Pyroptosis Of Osteoblasts Infected With Brucella And Inhibitory Effect Of Melatonin On Pyroptosis

Objective:To verify whether Brucella can induce pyroptosis in osteoblasts,and evaluate the anti-pyroptosis effect of melatonin on Brucella infection on cell level.In order to establishing a theoretical foundation for melatonin treatment of brucellosis spondylitis.Methods:(1)Establishing the model of Brucella infection of osteoblasts by incubating Brucella with osteoblasts.(2)Total proteins were extracted from osteoblasts after lysing.Utilizing western blot to detect whether Brucella induced the upregulation of pyroptosis related proteins in osteoblasts.Constructing osteoblast cell growing on the glass slides and incubating with Brucella.Morphological changes of pyroptosis of osteoblasts were observed by using scanning electron microscope(SEM)after fixation.(3)The effect of Brucella on osteoblast viability was detected by CCK-8 experiment.LDH assay was used to detect whether Brucella caused the membrane rupture and the cell contents release of osteoblasts.The anti-pyroptosis effect of melatonin on Brucella infection of osteoblasts was evaluated by CCK-8 experiment,LDH assay and western blot.Results:(1)Brucella could invade osteoblasts and multiply in the cells which reflected that the cell model was successfully constructed.(2)The stripes and data of western blot showed that Brucella infection induced up-regulated expression of pyroptosis related proteins in osteoblasts.The morphological changes of osteoblasts infected by Brucella were observed by using SEM,which proved that Brucella infection caused the pyroptosis of osteoblasts.(3)The results of CCK-8 experiment showed that the viability of osteoblasts decreased due to Brucella infection.The results of LDH assay showed that Brucella infection induced the membrane rupture and cell contents release of osteoblasts.The above results comprehensively proved that Brucella infection caused the decrease of osteoblast viability and induced pyroptosis thereof.By contrast,melatonin treatment could significantly alleviate the above changes.Conclusions:Brucella infection induces the decrease of osteoblast viability and induces pyroptosis thereof.Melatonin has a good anti-pyroptosis effect on osteoblasts caused by Brucella infection,which can provide a new idea for the treatment of brucellosis spondylitis.