RESUMO
PURPOSE: We aim to establish an LPS-induced human aortic endothelial cells (HAECs) inflammatory injury model and explore the optimal conditions for inducing its injury. We expect to provide modeling references for the related experiments of vascular inflammatory diseases. METHODS: HAECs were cultured in vitro and treated with different concentrations of lipopolysaccharide (LPS) (0.1, 1, 10, 50, 100⯵g/mL) for 6, 12, and 24â¯h to establish the HAECs inflammatory injury model. The cell viability was determined by CCK-8 assay; the expression levels of inflammatory cytokines in the cells were detected by RT-PCRï¼the apoptosis rate of the cells was detected by flow cytometry. RESULTS: â Within 24â¯h of LPS treatment, the cell viability of the 0.1 and 1⯵g/mL groups showed an overall increasing trend with time, while the cell viability of the 10, 50, and 100⯵g/mL groups increased first and then decreased with time, and the cell viability of 50 and 100⯵g/mL groups was significantly lower than the normal control group at 24â¯h (P<0.01). â¡ RT-PCR results showed that after 50 and 100⯵g/mL LPS for 24â¯h, the inflammatory cytokines all showed an apparent upward trend compared with the normal control group (P<0.05), which was more significant in the 100⯵g/mL group. ⢠After 100⯵g/mL LPS for 24â¯h, the apoptotic necrosis rate of HAECs was higher than the normal control group (P<0.01). CONCLUSIONS: This experiment successfully established a HAECs injury model, indicating that the optimal conditions for inducing injury are an LPS concentration of 100⯵g/mL and a treatment time of 24â¯h.