当归饮子抑制IL-33介导的肥大细胞脱颗粒缓解荨麻疹样病变小鼠过敏反应的效应机制  

Effect of Danggui Yinzi(当归饮子)on Relieving Allergic Reactions in UL Mice by Inhibiting IL-33-mediated Degranulation of Mast Cells

作　　者：张晓桐 魏芹 蔡琛 庞尧斌 何影洁 吴石 郭静[1] 曾进浩 ZHANG Xiaotong;WEI Qin;CAI Chen;PANG Yaobin;HE Yingjie;WU Shi;GUO Jing;ZENG Jinhao(Hospital of Chengdu University of Traditional Chinese Medicine,Chengdu 610072,Sichuan,China;Ordos Hospital of Traditional Chinese Medicine,Ordos 017010,Inner Mongolia,China)

机构地区：[1]成都中医药大学附属医院,四川成都610072 [2]鄂尔多斯市中医医院,内蒙古鄂尔多斯017010

出　　处：《中华中医药学刊》2021年第11期144-148,I0022,I0023,I0024,共8页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金(81873310);中国博士后科学基金(2019M651560);四川省科学技术厅科技计划(2018JY0660);成都中医药大学“杏林学者”学科人才研究专项(030041139)。

摘　　要：目的研究当归饮子对荨麻疹样病变(UL)小鼠皮肤肥大细胞活化、脱颗粒反应的影响及IL-33介导的细胞因子TNF-α、IL-1β的干预机制。方法雄性BALB/c小鼠随机分为空白对照组(生理盐水20 mL·kg^(-1)·d)、模型对照组(生理盐水20 mL·kg^(-1)·d)、氯雷他定组(0.0013 g·kg^(-1)·d)、当归饮子高、中、低剂量组(39.3、19.6、9.8 g·kg^(-1)·d)。造模:运用腹腔注射卵白蛋白(10μg)与氢氧化铝悬液(100μL)进行致敏并激发复制UL过敏反应小鼠模型。检测:苏木素-伊红(HE)染色观察小鼠皮肤组织病理学改变;甲苯胺蓝染色观察小鼠皮肤组织肥大细胞活化及脱颗粒现象;免疫组化(IHC)法检测皮肤组织肥大细胞脱颗粒中类胰蛋白酶(MCT)蛋白的表达;酶联免疫吸附法(Elisa)检测血清中TNF-α、IL-1β水平;采用实时荧光定量聚合酶链式反应(Real-time PCR)法检测皮肤组织IL-33 mRNA转录水平。结果当归饮子可显著改善UL小鼠皮肤真皮水肿、胶原纤维断裂、毛细血管扩张等病理表现;并能减轻UL小鼠皮肤组织肥大细胞的活化、脱颗粒反应,抑制肥大细胞脱颗粒中MCT蛋白的表达(P<0.05)。与空白对照组相比,模型对照组UL小鼠血清中TNF-α、IL-1β水平显著升高(P<0.01);与模型对照组相比,当归饮子能显著下调UL小鼠血清中TNF-α、IL-1β水平,以高剂量组疗效较好(P<0.01)。与空白对照组比较,模型对照组UL小鼠皮肤组织的IL-33 mRNA表达水平显著增加(P<0.01);与模型对照组比较,当归饮子能显著下调UL小鼠皮肤组织的IL-33 mRNA表达水平(P<0.01)。结论当归饮子可显著抑制UL小鼠皮肤组织肥大细胞活化、脱颗粒反应,改善皮肤真皮水肿、胶原纤维断裂等病理表现,其机制可能与其下调IL-33介导的细胞因子TNF-α、IL-1β的分泌有关。Objective To investigate the effect of Danggui Yinzi(当归饮子)on the activation and degranulation of skin mast cells in urticaria-like lesions(UL)mice and the intervention mechanism of secretion of TNF-αand IL-1βmediated by IL-33.Method Grouping:Male BALB/c mice were randomly divided into blank group(normal saline 20 mL·kg^(-1)·d),model group(normal saline 20 mL·kg^(-1)·d),loratadine group(0.0013 g·kg^(-1)·d)and high,medium and low dose groups of Danggui Yinzi(39.3,19.6,9.8 g·kg^(-1)·d).Modeling:To develop urticarial lesions,the animals,except normal controls,were sensitized intraperitoneally(i.p.)with 1 mL aluminum hydroxide suspension(alum,dissolved in 0.9%NaCl,10 g/L)containing 1 mg of ovalbumin.Histopathological changes of mouce skin was observed by hematoxylin-eosin(HE)staining.The degranulation of mast cells in mouse skin was determined by toluidine blue staining.The mast cell tryptase expression in skin tissue was detected by immunohistochemistry(IHC).The serum tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)levels were detected by enzyme-linked immunosorbent assay(ELISA).The Interleukin-33(IL-33)mRNA transcription levels in skin tissue were detected by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result Danggui Yinzi can significantly improve the pathological manifestations of UL mouse dermal edema,collagen bundle separation,telangiectasia and can reduce the activation and degranulation reaction of mast cells in the skin tissue of UL mice,and inhibit the MCT protein in the degranulation of mast cells expression(P<0.05).Compared with those of the blank group,the serum levels of TNF-αand IL-1βin UL mice in the model group were significantly increased(P<0.01).Compared with the model group,Danggui Yinzi could significantly down-regulate the levels of TNF-αand IL-1βin UL mice’s serum and the highest dose group had the best effect(P<0.01).Compared with that of the blank group,the IL-33 mRNA expression level in the skin tissue of UL mice in the model

关 键 词：荨麻疹样病变 肥大细胞脱颗粒 当归饮子 肥大细胞类胰蛋白酶 白细胞介素33 

分 类 号：R285.5[医药卫生—中药学]