变应性鼻炎(AR)是与免疫相关，以组胺为主要效应分子的常见病，反复发作难以根治，西医治疗的最大困难是远期疗效欠满意。源于《伤寒杂病论》的小青龙汤治疗AR具有可长期用药和远期效果良好的特点。研究表明其具有调节免疫和抗组胺的双重作用，这恰好与AR的病理过程直接相关，然而，小青龙汤在AR中调节免疫和抗组胺作用的机制及通路尚不明晰。最新研究表明组胺通过PKCδ/ERK/PARP-1信号通路促进H1R基因表达增强。因此，本项目采用OVA致敏AR豚鼠模型，以及RBL-2H3和富含H1R的Hela细胞株，检测小青龙汤对鼻黏膜Th1/Th2细胞因子、H1R、组氨酸脱羧酶（HDC）表达，对H1R蛋白浓度、HDC活性、组胺含量，对肥大细胞脱颗粒释放组胺酸，以及对与H1R表达密切相关的PKCδ/ERK/PARP-1信号通路的影响，探讨小青龙汤治疗AR的药理机制，为其临床推广应用和进一步的开发提供研究基础。

Allergic Rhinitis(AR) is a common disease which is related to allergy reaction and histamine is the main effect molecule. AR is difficult to be cured and often recurrent attacks. The largest difficulty of west treatment is poor long-term outcome. Xiao-Qing-Long-Tang, comes from Shang-han Za-bing Lun, has been used to treat allergic rhinitis against allergy in clinic. It regulates Th1/Th2 cytokines and antagonize histamine. We investigated the effect of Xiao-Qing-Long-Tang on ovalbumin(OVA)-induced allergic behavior, specific IgE in serum, the mRNA levels of Th1/Th2 cytokine genes closely related to histamine signaling in guinea pig nasal mucosa. We also investigated the effect of Xiao-Qing-Long-Tang on OVA provocation caused acute allergic symptoms accompanied with up-regulations of H1R and HDC mRNAs and increases in HDC activity, histamine content in the nasal mucosa, and related histamine signaling including mRNA levels of histamine H1 receptor (H1R) and histidine decarboxylase (HDC), H1R and HDC activities, and histamine content in guinea pig nasal mucosa. We also investigated the effect of Xiao-Qing-Long-Tang on the degranulation of mast cell. Recently, we demonstrated that histamine stimulation induced the upregulation of H1R gene expression through the protein kinase Cδ (PKCδ)/extracellular signal-regulated kinase/poly(ADP-ribose) polymerase-1 signaling pathway in HeLa cells expressing H1R endogenously. So we examined the effect of Xiao Qing Long Tang on histamine induced up-regulation of H1R gene expression via PKCδ/ERK/PARP-1 signaling pathway in HeLa cells. We discuss the pharmacological mechanism of Xiao-Qing-Long-Tang treating AR. To promote clinical popularization and application of Xiao-Qing-Long-Tang, and provide study basis for its further development.

变应性鼻炎（AR）的治疗可通过抗过敏与抗炎途径实现。本项目采用OVA致敏AR豚鼠、小鼠模型，以及RBL-2H3、RAW264.7和HNECs细胞株，检测小青龙汤对鼻黏膜HDC、H1R mRNA表达和H1R蛋白浓度，血清Th1/Th2细胞因子、组胺含量，肥大细胞脱颗粒释放HDC，PKCδ/ERK/PARP-1信号通路和TLR4/NF-κB炎症信号通路的影响；采用高效液相-质谱联用仪检测小青龙汤含药血清，确定其成分，即小青龙汤发挥作用可能的物质基础；并通过组胺诱导豚鼠离体回肠收缩实验确定小青龙汤中拮抗组胺的有效成分，利用LPS诱导的RAW264.7和组胺诱导的HNECs细胞炎症模型确定其抗炎作用的有效成分。 结果：1.与AR模型组比较，小青龙汤可显著降低血清IL-4、IL-5、IL-13、组胺、特异性IgE水平，不能调节血清IFN-γ水平。2.小青龙汤可降低HDC mRNA表达，抑制肥大细胞脱颗粒，但不通过降低H1R mRNA表达降低H1R蛋白浓度。3.小青龙汤及其有效成分对PKCδ/ERK/PARP-1信号通路作用不显著，但可通过抑制TLR4/NF-κB炎症信号通路降低IL-6、IL-10、TNF-α、NO炎症因子，抑制巨噬细胞活化及其吞噬功能。4.经HPLC-MS检测得到小青龙汤入血成分含有麻黄碱、肉桂酸、儿茶素、五味子醇甲、6-姜辣素、甘草苷、甘草酸、芍药苷。其中，儿茶素、麻黄碱可使组胺量效曲线向右下方移动，具有拮抗组胺H1受体的作用。儿茶素、五味子醇甲可抑制LPS诱导的RAW264.7细胞释放NO、IL-6等炎症因子分泌，降低巨噬细胞表面CD86、MHC II表达，以及减弱巨噬细胞吞噬功能，抑制胞浆中iNOS表达、细胞膜上TLR4表达，以及胞核NF-κBp65表达。 结论：1.小青龙汤可通过降低血清Th2细胞因子、特异性IgE水平调整AR的Th1/Th2免疫失衡；2.小青龙汤可降低HDC mRNA表达而降低组胺含量，抑制肥大细胞脱颗粒，以H1受体的非竞争性拮抗剂形式发挥抗组胺作用；3.小青龙汤通过抑制TLR4/NF-κB炎症信号通路起到抗炎作用。4.小青龙汤入血成分麻黄碱、儿茶素具有抗组胺作用，儿茶素、五味子醇甲具有抗炎作用，抗组胺与抗炎起协同作用。本项目探讨了小青龙汤治疗AR疗效机制，并分析出其有效单体，为其临床推广和进一步的开发提供研究基础。