基于两模两方模式的小青龙汤、麻黄附子细辛汤对两种不同AR模型干预机制的方证相应研究

Double formulae for double patterns serve as inevitable approach for further study of formula pattern correspondence. This approach may shed light on the correlation between formula, medicinal, disease and pattern. This approach may also represent the latest innovative trend in integrating traditional Chinese medicine thinking and modern scientific technology. The research studied subjects with allergic rhinitis (AR) by observing their alertness, aversion to cold, body temperature, weight increase, blood cAMP/cGMP, kidney and testes morphology to determine yang deficiency pattern. Nasal allergy, PCA, histamine provocation test, and eosinophil rate were measured for immune response evaluation. Technologies like ELISA, Flow Cytometry, RT-PCR were employed to observe if there were any abnormal drift of Th1/Th2 cell subtype. Therefore three different yang deficiency patterns in AR guinea pigs at tissue, cellular and molecular levels were evaluated from both pathogenic and antipathogenic perspectives. Xi?o Qīng Lóng Tāng (Minor Green Dragon Decoction) for dissipating cold and Má Huáng Fù Z? Xì Xīn Tāng (Ephedra, Aconite and Asarum Decoction) for warming meridians and dissipating cold were crosstabed in usage to treat AR subjects of different patterns. This research aims to screen and establish a reliable animal model of yang deficiency pattern with Allergic Rhinitis by evaluating its efficacy for patterns, which will provide a paradigm for experimental studies of double formula for double patterns using animal models.

"两模两方"是深化方证相应研究的必然要求。这种研究思路有助于揭示方药与病证之间的关联度大小，能够突出中医学思维特征与现代科学设计融合的特色，是方证关系研究的最新实验模式。本项目拟以变应性鼻炎（AR）为切入点，通过观察动物反应灵敏程度、畏寒、体温、体重增长、血浆cAMP/cGMP含量和肾、睾丸组织学变化来判断各组肾阳虚程度，根据鼻部过敏症状、PCA试验、组胺刺激试验、嗜酸细胞阳性率等判断免疫应答反应程度，采用ELISA、流式细胞术、RT-PCR等技术观察Th1/Th2细胞亚型异常漂移，在组织、细胞和分子水平上从正邪两方对三种不同的阳虚复合性AR豚鼠模型进行评价，用单纯散寒的小青龙汤与温经散寒的麻黄附子细辛汤对单纯性和复合性两种病机有差异的AR模型进行交叉干预。本项目旨在筛选、建立稳定的阳虚型AR复合动物模型，对病机与治法一致与相反情况下出现的客观疗效进行验证，以便为两模两方实验研究提供范例。

本项目严格按照计划进行，基本完成原计划任务，达到预期目标：建立了OVA变应性鼻炎单纯致敏豚鼠模型，从三种肾阳虚诱导剂筛选确立了羟甲脲所致肾阳虚型复合模型，比较分析了单纯模型和复合模型的病机属性。两模两方交叉干预，探讨小青龙汤和麻黄附子细辛汤分别对两种不同变应性鼻炎模型的疗效和作用机制。.结果显示，单纯模型和复合模型血清IFN-γ、脾细胞IFN-γmRNA、特异性转录因子T-bet-mRNA降低，而IL-4、IL-4-mRNA、特异性转录因子GATA-3 mRNA升高，IFN-γ/IL-4 、IFN-γmRNA/IL-4mRNA、T-bet mRNA/GATA-3 mRNA降低，细胞因子含量与相应mRNA的表达一致，提示单纯模型和复合模型均发生Th1/Th2失衡产生TH2型免疫应答。.小青龙汤可使单纯模型组IFN-γ、T-bet mRNA升高，使两种模型IL-4、GATA-3mRNA降低，IFN-γ/IL-4、T-bet mRNA/GATA-3mRNA回升；对复合模型IFN-γ、T-bet mRNA不影响；使两种模型IFN-γmRNA升高，IL-4mRNA降低，IFN-γmRNA/IL-4 mRNA升高，其作用在单纯模型组更明显。这表明，小青龙汤调节Th1/Th2失衡在单纯模型优于在复合模型，可提示其在外散风寒方面的优势。.麻黄附子细辛汤可使两种模型IFN-γ、T-bet mRNA升高，IL-4、GATA-3 mRNA降低，IFN-γ/IL-4、T-bet mRNA/GATA-3 mRNA升高，对复合模型组更明显；使复合模型组IFN-γmRNA升高，两种模型IL-4 mRNA降低，IFN-γmRNA/IL-4 mRNA升高；对单纯模型IFN-γmRNA无影响。因此，麻黄细辛附子汤调节Th1/Th2平衡在复合模型优于比在单纯模型，可提示在温阳方面的优势。.本项目验证了小青龙汤对单纯致敏模型的疗效，麻黄附子细辛汤对肾阳虚复合模型的疗效，以及在单纯致敏模型中小青龙汤优于麻黄细辛附子汤，在肾阳虚复合模型中麻黄细辛附子汤优于小青龙汤，诠释了方证相应的科学性，可为两模两方类研究提供借鉴。证明了该两首经方对同一现代疾病的客观疗效，有助于推动变应性鼻炎临床治疗水平；小青龙汤、麻黄细辛附子汤可以纠正Th1/Th2失衡，发挥多层面、多维度的调节作用，可以启发两方的现代药理机制研究。