首页|根尖牙乳头干细胞外泌体通过AHR调节CDC42转录促进血管内皮细胞迁移的机制研究

根尖牙乳头干细胞外泌体通过AHR调节CDC42转录促进血管内皮细胞迁移的机制研究

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目的:探究根尖牙乳头干细胞来源的外泌体(SCAP-Exo)调控细胞分裂周期蛋白42(CDC42)转录介导血管内皮细胞(VECs)迁移的分子机制.方法:超速离心法提取SCAP-Exo并应用透射电镜和纳米粒子追踪实验鉴定.构建CDC42低表达的VECs,应用SCAP-Exo处理VECs,实时荧光定量PCR与Western blot实验检测SCAP-Exo处理后CDC42的基因和蛋白表达水平,通过划痕实验和Transwell迁移实验检测VECs迁移能力.通过Western blot和免疫荧光染色实验,检测SCAP-Exo对VECs中芳香烃受体(AHR)核转位以及CDC42表达的影响.结果:SCAP-Exo可以上调CDC42基因和蛋白表达水平,增强VECs的迁移能力.与SCAP-Exo处理的正常VECs相比,VECs低表达CDC42后加入SCAP-Exo,迁移能力明显降低.SCAP-Exo可以促进VECs内AHR转位至细胞核内,当抑制VECs中AHR核转位后,VECs内CDC42的基因和蛋白表达水平下降.结论:SCAP-Exo通过激活VECs内AHR入核促进CDC42转录,进而上调CDC42的蛋白表达水平,从而增强VECs的迁移能力.
Exosomes derived from stem cells from apical papilla promotes the migration of vascular endothelial cells by regulating CDC42 transcription via AHR
Objective:To explore the potential mechanism by which exosomes derived from stem cells from apical papilla(SCAP-Exo)regulate the migration of vascular endothelial cells(VECs)by targeting cell division cycle 42(CDC42)transcription.Methods:SCAP-Exo extracted through ultracentrifugation were characterized by transmission electron microscopy and nanoparticle tracking analy-sis.VECs transfected with siCDC42 were treated with SCAP-Exo,and real-time quantitative PCR and western blot assays were conduc-ted to detect the mRNA and protein expression levels of CDC42 after SCAP-Exo treatment.Scratch wound healing and transwell cell mi-gration assays were performed to assess the migration of VECs.The effects of SCAP-Exo on aryl hydrocarbon receptor(AHR)nuclear translocation and CDC42 expression in VECs were detected by western blotting and immunofluorescence staining assays.Results:SCAP-Exo could upregulate the mRNA and protein expression levels of CDC42 and enhance the migration of VECs compared to those in the Control group.The migration rate of VECs were reduced when VECs with siCDC42 were treated with SCAP-Exo compared to the SCAP-Exo-treated VECs.SCAP-Exo induced AHR translocation to the nucleus in VECs.After inhibiting AHR nuclear translocation in VECs,the mRNA and protein expression levels of CDC42 decreased.Conclusions:SCAP-Exo promoted CDC42 transcription by acti-vating AHR translocation,thereby upregulating CDC42 expression,which promoted the migration of VECs.

stem cells from apical papillaexosomevascular endothelial cellcell migrationcell division cycle 42aryl hydro-carbon receptor

马元、庄雪莹、祝雪松、刘尧、陈旭

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中国医科大学口腔医学院·附属口腔医院儿童口腔科,辽宁省口腔疾病重点实验室,辽宁沈阳 110002

根尖牙乳头干细胞 外泌体 血管内皮细胞 细胞迁移 细胞分裂周期蛋白42 芳香烃受体

2024

口腔生物医学
南京医科大学

口腔生物医学

CSTPCD
影响因子:0.423
ISSN:1674-8603
年,卷(期):2024.15(6)