基于NF-κB/NLRP3/IL-1β信号通路探讨荆防合剂对荨麻疹小鼠干预作用的研究  

Intervention effect of Jingfang Mixture on urticaria mice based on NF-κB/NLRP3/IL-1βsignaling pathway

作　　者：李市荣 王西双 程国良 孙成宏 李艳芳 岳茹婧 曾振 姚景春 LI Shi-rong;WANG Xi-shuang;CHENG Guo-liang;SUN Cheng-hong;LI Yan-fang;YUE Ru-jing;ZENG Zhen;YAO Jing-chun(Center for New Drug Pharmacology,Lunan Pharmaceutical Group Co.,Ltd.,Linyi 273400,China;State Key Laboratory of Generic Manufacture Technology of Chinese Traditional Medicine,Linyi 276006,China;School of Traditional Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]鲁南制药集团股份有限公司新药药理中心,山东临沂273400 [2]中药制药共性技术国家重点实验室,山东临沂276006 [3]北京中医药大学中医学院,北京100029

出　　处：《中国中药杂志》2022年第20期5467-5472,共6页China Journal of Chinese Materia Medica

基　　金：山东省重大科技创新工程项目(2021CXGC010508);临沂市2019年科技创新重大专项(2019ZDZX001)。

摘　　要：观察荆防合剂对氢氧化铝/卵白蛋白诱导的荨麻疹小鼠的疗效并探讨其作用机制。将60只雄性昆明小鼠随机均分为正常组,模型组,荆防合剂低、中、高剂量组,盐酸西替利嗪片阳性药组。腹腔注射卵白蛋白与氢氧化铝混合液建立荨麻疹小鼠模型。第2次免疫时观察并记录小鼠瘙痒情况;苏木精-伊红(HE)染色法检测各组小鼠皮肤组织病理变化;酶联免疫吸附测定(ELISA)法检测各组小鼠血清中白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)的水平;免疫组织化学法检测各组小鼠皮肤组织中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、IL-1β的表达;蛋白免疫印迹(Western blot)法检测各组小鼠皮肤组织中核因子κB(NF-κB p65)、NLRP3、凋亡相关微粒蛋白(ASC)、胱冬肽酶-1(caspase-1)、IL-1β蛋白的表达水平。结果显示,除正常组外,荆防合剂低、中、高剂量组,阳性药组以及模型组小鼠均出现不同程度的瘙痒反应。与模型组比较,荆防合剂低、中、高剂量组以及阳性药组小鼠瘙抓潜伏期延长(P<0.05),瘙抓次数明显减少(P<0.05);皮肤组织病理形态显著改善;血清中IL-1β和TNF-α的水平显著降低(P<0.05);皮肤组织中NLRP3和IL-1β阳性表达细胞数量显著减少(P<0.01);p-NF-κB p65、NLRP3、ASC、cleaved caspase-1、IL-1β蛋白的表达显著下调(P<0.05)。综上研究结果表明,荆防合剂能够抑制荨麻疹小鼠炎症反应,其机制可能与抑制NF-κB/NLRP3/IL-1β信号通路激活有关。This study explored the curative effect of Jingfang Mixture on urticaria mice induced by aluminum hydroxide/ovalbumin,and discussed its mechanism.Sixty male Kunming mice were randomly divided into a normal group,a model group,three Jingfang Mixture(low-dose,medium-dose,and high-dose)groups,and a positive drug(cetirizine hydrochloride)group.The urticarial model in mice was induced by the intraperitoneal injection of the mixed solution of ovalbumin and aluminum hydroxide.The degrees of pruritus were observed after the second immunization.Pathological changes were detected by hematoxylin and eosin(HE)staining.Levels of interleukin 1β(IL-1β)and tumor necrosis factorα(TNF-α)in the serum were detected by enzyme linked immunosorbent assay(ELISA).Expressions of NOD-like receptor protein 3(NLRP3)and IL-1βwere detected by immunohistochemistry(IHC).Expressions of nuclear factor kappa-B(NF-κB p65),NLRP3,apoptosis-associated speck-like protein containing a CARD(ASC),cysteinyl aspartate-specific proteases 1(caspase-1),and IL-1βproteins were detected by Western blot.The results showed that,except for the normal group,the mice in all groups had different degrees of pruritus.Compared with the model group,the Jingfang Mixture groups and the positive drug group prolonged the scratching latency of mice(P<0.05),and significantly reduced the number of scratching(P<0.05).In addition,the Jingfang Mixture groups and the positive drug group improved the pathological morphology of skin tissue.The expression levels of IL-1βand TNF-αin serum were significantly reduced(P<0.05),and the number of NLRP3 and IL-1βpositive cells was decreased(P<0.01).The expressions of p-NF-κB p65,NLRP3,ASC,cleaved caspase-1,and IL-1βprotein were significantly down-regulated(P<0.05).The results of the above study indicate that Jingfang Mixture inhibit the inflammatory response in urticaria mice,and the mechanism may be related to the inhibition of activating NF-κB/NLRP3/IL-1βsignaling pathway.

关 键 词：荆防合剂 荨麻疹小鼠 NF-ΚB NLRP3 IL-1Β 炎症 

分 类 号：R285.5[医药卫生—中药学]