Effect of lipopolysaccharide-induced central nervous system inflammation response on blood-brain barrier function in vitro

Objective To establish a blood-brain barrier (BBB) model in vitro using bEND.3 mouse brain microvascular endothelial cells and astrocytes,and observe the effects of central nervous system inflammation induced by lipopolysaccharide (LPS) on blood-brain barrier function. Methods The primary brain microvascular astrocytes of mice were cultured,and their cell purity was identified by flow cytometry using glial fibrillary acidic protein (GFAP) as marker.The primary astrocyte was co-cultured with bEND.3 mouse brain microvascular endothelial cells to establish an in vitro blood-brain barrier model.Then the primary astrocytes,bEND.3 brain microvascular endothelial cells,and co-cultured blood-brain barrier models were divided into control group and LPS treatment group.The LPS treatment group was stimulated with LPS (20μg / ml) for 24 hours.The blood-brain barrier function was measured by trans-epithelium electrical resistant (TEER),and western blot was used to measure the relative expression of Occludin and ZO-1 protein. Results The primary cultured astrocytes had a typical polymorphic morphology,and the purity of the cells identified by flow cytometry was 96%.After treated with LPS,the TEER values of three types of cells were significantly lower than those of the control group (P<0.05),and the relative expression levels of Occludin and ZO-1 proteins were also significantly lower than those of the control group (P<0.05). Conclusion Co-culture of bEND.3 mouse brain microvascular endothelial cells and astrocytes can successfully establish an in vitro blood-brain barrier model.LPS-induced central nervous system inflammation can impair the integrity of the blood-brain barrier and destroy its function.