实用医学杂志2025，Vol.41Issue(21)：3345-3351,7.DOI:10.3969/j.issn.1006-5725.2025.21.007
维生素B12通过下调自噬增强HDM处理的人气道上皮细胞中ZO-1的表达
Vitamin B12 enhances ZO-1 expression in HDM-treated human airway epithelial cells by down-regulating autophagy
摘要
Abstract
Objective To investigate the effect of vitamin B12(VB12)on the expression of zonula occludens-1(ZO-1)in house dust mite(HDM)-treated human airway epithelial cell line(Beas-2b)and its underlying mechanism.Methods Beas-2b cells were cultured in DMEM high-glucose medium containing 10%fetal bovine serum.The cells were divided into four groups:control,VB12,HDM,and VB12+HDM.Beas-2b cells were trans-fected with lentiviruses carrying NC-siRNA,ATG5-siRNA,BECN1-siRNA,and mCherry-EGFP-LC3.After 12 hours of transfection(MOI=20),the medium was replaced with fresh medium,and stable transfected cell lines were selected using puromycin(1 µg/mL).Cells were stimulated with VB12(20 µg/mL)and HDM(50 µg/mL)for 24 hours.The protein levels of ZO-1,autophagy-related protein 5(ATG5),BECN1 and microtubule-associated protein light chain 3(LC3)were detected by immunofluorescence and Western blot.Autophagy in human airway epithelial cells was observed using confocal microscopy.Results Compared with the control group,the expression of ZO-1 in the HDM group was lower(P<0.05),while the expressions of ATG5,BECN1,and LC3 were higher(P<0.05).Compared with the HDM group,the VB12+HDM group showed increased ZO-1 expression(P<0.05),decreased expressions of ATG5,BECN1,and LC3(P<0.01),and reduced autophagosome formation(P<0.05).In ATG5-and BECN1-knockdown cell lines,ZO-1 expression increased after HDM treatment(P<0.05).Conclusion Vb12 can enhance ZO-1 expression in HDM-treated human airway epithelial cells by down-regulating autophagy,and its mechanism is associated with the ATG5 and BECN1 signaling pathways.关键词
维生素B12/闭锁小带蛋白-1/自噬/自噬相关蛋白5/苄氯素1Key words
vitamin b12/zonula occludens-1/autophagy/ATG5/BECN1分类
医药卫生引用本文复制引用
李月蛟,蓝楠,王星,唐红梅,王志彬,张沄,袁谢芳,王孝芸..维生素B12通过下调自噬增强HDM处理的人气道上皮细胞中ZO-1的表达[J].实用医学杂志,2025,41(21):3345-3351,7.基金项目
国家自然科学基金项目(编号:82100021) （编号:82100021）
西南医科大学校级科研项目(编号:2023QN043) （编号:2023QN043）
