High expression of IL-1p in coronary artery endothelial cells promotes neutrophil activation and Sema4D secretion in Ka-wasaki disease
Objective To investigate the effects and mechanisms of human coronary artery endothelial cell(HCAEC)conditioned medium on the biological functions of neutrophils in Kawasaki disease(KD).Methods Pooled serum were collected from KD patients and healthy children in December 2022 in Xi'an Children's Hospital to stimulate HCAECs,named as KD group and HC group.RT-PCR was performed to detect the expression of IL-1 β in HCAECs in two groups.Additionally,HCAECs were divided into si-IL-1 βgroup and si-NC group,and respectively transfected with IL-1 β siRNA and invalid siRNA,and then stimulated with KD pooled serum.RT-PCR was used to measure the mRNA expression of IL-1 β in si-IL-1 β group and si-NC group.The supernatants in both si-IL-1 βgroup and si-NC group were collected after centrifugation and filtration to obtain the conditioned medium.The concentration of IL-1 β in the conditioned medium was measured by ELISA.Transwell experiment was performed to assess the effects of the conditioned medium on neutrophil migration.Flow cytometry was used to assess the effects of conditioned medium on neutrophil apoptosis and surface expression of Sema4D.Neutrophils were pretreated with the metalloproteinase ADAM17 inhibitor TAPI-1 or DMSO,and stimulated with the conditioned medium from si-IL-1 β group or si-NC group,namely TAPI-1 pretreatment+si-NC conditioned medium group,DMSO pretreatment+si-NC conditioned medium group,TAPI-1 pretreatment+si-IL-1 β conditioned medium group and DMSO pretreatment+si-IL-1 β conditioned medium group.Flow cytometry was used to measure Sema4D expression in neutrophils.Results RT-PCR showed that the expression of IL-1 β in HCAECs in KD group was significantly higher than that in HC group(P＜0.01),and the IL-1 βmRNA expression in si-IL-1 β group was significantly lower than that in si-NC group(P＜0.01).Compared to si-NC group,the IL-1 βconcentration in conditioned medium was significantly decreased in si-IL-1β group(P＜0.01),the migration ability of neutrophils was also significantly reduced(P＜0.01),while the neutrophil apoptosis and the Sema4D expression were increased(P＜0.01).Compared to DMSO pretreatment+si-NC conditioned medium group,the percentage of Sema4D+neutrophils was significantly increased in TAPI-1 pretreatment+si-NC conditioned medium group(P＜0.05),while there was no significant difference in neutrophil Sema4D expression between TAPI-1 pretreatment+si-IL-1β conditioned medium group and DMSO pretreatment+si-IL-1 β conditioned medium group(P＞0.05).Conclusion Increased expression of IL-1 β in endothelial cells promotes the neutrophil migration,and inhibits the neutrophil apoptosis,and may enhance the secretion of Sema4D by neutrophils through ADAM17 activation in Kawasaki disease.
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维普
