摘要
目的研究布地奈德(BUD)对S100钙结合蛋白A4(S100A4)诱导的肥大细胞活化、炎性因子释放及受体表达的影响。方法8~10周龄野生型(WT)的C57BL/6健康雄性小鼠2只,取胫骨与股骨提取骨髓进行骨髓源肥大细胞(BMMCs)培养,将5 mL PBS注射至小鼠腹腔,取腹腔灌洗液以获取腹膜来源的肥大细胞(PMCs);采用S100A4蛋白与BUD处理小鼠BMMCs和PMCs,实验分为PBS组、S100A4组、BUD组及S100A4+BUD组;β-己糖胺酶(β-hex)释放实验检测细胞脱颗粒指标β-hex、ELISA测定细胞活化介质类胰蛋白酶、糜蛋白酶及白三烯B4的释放,流式细胞术编码微球芯片技术(CBA)法检测炎性因子(IL-5、IL-6、IL-13和TNF-α)分泌水平以及Toll样受体4(TLR4)和晚期糖基化终产物受体(RAGE)的表达。结果在BMMCs与PMCs中,与PBS组相比,S100A4组培养上清中β-hex、类胰蛋白酶、白三烯B4及相关炎性因子IL-5、IL-6、IL-13和TNF-α的释放增加,细胞上TLR4和RAGE的表达上调(P<0.05);与S100A4组相比,S100A4+BUD组肥大细胞活化及炎性因子分泌明显被抑制,同时TLR4的表达下调(P<0.05),而RAGE的表达无显著变化(P>0.05)。结论BUD能够抑制S100A4介导的肥大细胞活化及炎性因子释放,并下调TLR4的表达,但不影响RAGE的表达,BUD可能通过TLR-4受体发挥其功能,为BUD在过敏性炎症中的应用提供新的理论依据。
Objective To investigate the effects of budesonide(BUD)on S100A4-induced mast cell activation,release of inflammatory factors,and receptor expression.Methods Bone marrow was extracted from the tibiae and femurs of two 8-10-week-old wild-type(WT)C57BL/6 healthy male mice to culture bone marrow-derived mast cells(BMMCs).Peritoneal lavage was performed by injecting 5 mL of PBS into the mouse peritoneal cavity to obtain peritoneal mast cells(PMCs).Mouse BMMCs and PMCs were treated with S100A4 protein and BUD.The experiment was divided into PBS group,S100A4 group,BUD group,and S100A4+BUD group.Theβ-hexosaminidase(β-hex)release assay was used to measure the degranulation markerβ-hex.ELISA was used to determine the release of activation mediators tryptase,chymase,and leukotriene B4.Cytokine bead array(CBA)flow cytometry was used to detect the secretion levels of inflammatory factors(IL-5,IL-6,IL-13,and TNF-α)and the expression of Toll-like receptor 4(TLR4)and receptor for advanced glycation end products(RAGE).Results In both BMMCs and PMCs,compared with the PBS group,the S100A4 group showed increased release ofβ-hex,tryptase,leukotriene B4,and the related inflammatory factors IL-5,IL-6,IL-13,and TNF-αin the culture supernatant,as well as upregulated expression of TLR4 and RAGE on the cells(P<0.05).Compared with the S100A4 group,the S100A4+BUD group showed significant inhibition of mast cell activation and inflammatory factor secretion,along with downregulated expression of TLR4(P<0.05),while the expression of RAGE showed no significant change(P>0.05).Conclusion BUD can inhibit S100A4-mediated mast cell activation and inflammatory factor release,and downregulate the expression of TLR4,but does not affect the expression of RAGE.BUD may exert its function through the TLR-4 receptor,providing a new theoretical basis for the application of BUD in allergic inflammation.
作者
陶甜
杨茂婕
潘忍
吴通前
何菁菁
晏世荣
符竹
王颜香
张廷香
张天霞
杨俊
余芳
TAO Tian;YANG Maojie;PAN Ren;WU Tongqian;HE Jingjing;YAN Shirong;FU Zhu;WANG Yanxiang;ZHANG Tingxiang;ZHANG Tianxia;YANG Jun;YU Fang(School of Medical Laboratory,Guizhou Medical University,Guiyang 550004,Guizhou,China;Clinical Research Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Departments of Clinical Laboratory,Guizhou Hospital of the First Affiliated Hospital of Sun Yat-sen University,Guiyang 550004,Guizhou,China;Clinical Laboratory Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China;Children Medical Center,the Affiliated Hospital of Guizhou Medical University,Guiyang 550004,Guizhou,China)
出处
《贵州医科大学学报》
2026年第1期49-56,共8页
Journal of Guizhou Medical University
基金
国家自然科学基金项目(82260324,82201968)
贵州省基础研究计划项目(黔科合基础-ZK[2023]一般394,黔科合基础-ZK[2023]一般398,黔科合基础-ZK[2025]面上442)
贵州省卫健委科学技术基金项目(2025GZWJKJXM1401)
贵州省2023年大学生创新训练项目(S202310660012)。
