蓝萼甲素通过抑制HMGB1/TLR4/NF-κB信号通路减轻肥大细胞脱颗粒引起的过敏反应  

Glaucocalyxin A attenuates allergic responses by inhibiting mast cell degranulation through HMGB1/TLR4/NF-κB signaling pathways

作　　者：朴艺花[1,2] 宋艺兰 王知广 姜京植[2,3] 李莉[2,3] 徐畅[2,3] 朴颖[2,5] 朴红梅 延光海[2,3] PIAO Yi-hua;SONG Yi-lan;WANG Zhi-guang;JIANG Jing-zhi;LI Li;XU Chang;PIAO Ying;PIAO Hong-mei;YAN Guang-hai(Department of Critical Care Medicine,Hospital of Yanbian University,Yanji 133000,China;Jilin Key Laboratory for Immune and Targeting Research onCommonAllergic Diseases,Yanbian University,Yanji133002,China;Department of Anatomy,College of Medicine,Yanbian University,Yanji 133002,China;Respiratory and Critical Care Medicine,Hospital of Yanbian University,Yanji 133000,China;Department of Emergency Medicine,Hospital of Yanbian University,Yanji 133000,China)

机构地区：[1]延边大学附属医院重症医学科,吉林延吉133000 [2]延边大学,吉林省过敏性常见疾病免疫与靶向研究重点实验室,吉林延吉133002 [3]延边大学医学院解剖教研室,吉林延吉133002 [4]延边大学附属医院呼吸与危重症医学科,吉林延吉133000 [5]延边大学附属医院急诊内科学,吉林延吉133000

出　　处：《药学学报》2021年第1期201-207,共7页Acta Pharmaceutica Sinica

基　　金：国家自然科学基金资助项目(81970018,81860729).

摘　　要：研究蓝萼甲素(glaucocalyxin A,GLA)对肥大细胞介导的过敏反应的影响。动物福利和实验过程均遵循延边大学动物伦理委员会的规定。动物实验采用BALB/c小鼠耳部皮内注射anti-DNP-IgE(anti-dinitrophenylimmunoglobulin E)致敏,尾静脉注射DNP-HSA(human serum albumin)和4%伊文思蓝混合液激发制备动物皮肤被动过敏反应(passive cutaneous anaphylaxis,PCA)模型,收集双耳用于测量染料着色和组织学。细胞实验选用大鼠腹腔肥大细胞(rat peritoneal mast cells,RPMCs),分为对照组、IgE+antigen(Ag)组和IgE+Ag+GLA组,测定组胺释放以及钙内流水平。Western blot法检测高亲和力IgE受体(FcεRI)介导的信号通路蛋白及HMGB1/TLR4/NF-κB(high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B)信号蛋白。动物实验结果提示,GLA抑制小鼠PCA反应,减少伊文思蓝染料渗出,减轻耳朵炎症反应及耳朵厚度。细胞实验结果提示,GLA能减少组胺释放及钙离子内流,并抑制肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白介素(interleukin,IL)-4、IL-13和IL-1β生成;Western blot实验结果显示GLA可抑制FcεRI介导的脾酪氨酸激酶(spleen tyrosine kinase,Syk)、Lck/Yes新型酪氨酸激酶(Lck/Yes novel tyrosine kinase,Lyn)、酪氨酸激酶Fyn(tyrosine kinase,Fyn)、生长因子受体结合蛋白2(growth-factor receptor-bound protein 2,Gab2)和磷脂酶C-γ1(phospholipase C-γ1,PLCγ1)的磷酸化水平,同时GLA抑制HMGB1/TLR4信号通路使NF-κB p65核转移受限。结果表明,GLA通过HMGB1/TLR4/NF-κB信号通路抑制肥大细胞脱颗粒并减轻过敏性炎症。The study is to investigate the effect of glaucocalyxin A(GLA)on mast cell-mediated anaphylaxis.The animal welfare and experimental process of this experiment followed the regulations of the Animal Ethics Committee of Yanbian University.BALB/c mice were used in the animal experiment and randomly divided into five groups,control group,model group,and GLA low,medium,and high dose groups(10,20,and 40 mg·kg^(-1)).Mice were sensitized by intradermal injection of anti-dinitrophenyl-immunoglobulin E(DNP-IgE)into the ears and challenged with a mixture of DNP-human serum albumin(HSA)and 4%evans blue into the tail veins to prepare an animal skin passive cutaneous anaphylaxis(PCA)model,which was collected from both ears for measurement of dye staining and histology.Rat peritoneal mast cells(RPMCs)were used in the cell experiment and divided into control,IgE+antigen(Ag),and IgE+Ag+GLA groups to determine histamine release as well as calcium influx levels.High-affinity IgE receptor(FcεRI)-mediated signaling pathway proteins and HMGB1/TLR4/NF-κB(high mobility group box 1/toll like receptor 4/nuclear transcription factor kappa B)signaling proteins were detected by Western blot.The results of animal experiments suggest that GLA inhibits PCA,reduces evans blue dye exudation,and reduces ear inflammation and ear thickness in mice.The results of cellular experiments suggested that GLA could reduce histamine release and calcium influx,and inhibit tumor necrosis factor-α(TNF-α),interleukin(IL)-4,IL-13,and IL-1βproduction;Western blot results showed that GLA inhibited FcεRI-mediated phosphorylation levels of spleen tyrosine kinase(Syk),Lck/Yes novel tyrosine kinase(Lyn),tyrosine kinase Fyn(Fyn),growthfactor receptor-bound protein 2(Gab2),and phospholipase C(PLC)γ1,while GLA inhibited HMGB1/TLR4 signaling pathway to limit NF-κB p65 nuclear metastasis.The results indicate that GLA inhibits mast cell degranulation and attenuates allergic inflammation through the HMGB1/TLR4/NF-κB signaling pathway.

关 键 词：蓝萼甲素 高迁移率族蛋白B1 Toll样受体4 核转录因子-ΚB 肥大细胞脱颗粒 

分 类 号：R392.8[医药卫生—免疫学]