基于微透析-PK/PD相结合的“内病外治”自溶性微针的构建及其穴位给药作用机制研究

New form of acupoint administration was established and its corresponding mechanism was explored, which could provide a new idea and scientific basic for treatment of chronic asthrma. Therefore, in this study, drug loaded self-dissolving microneedle was prepared and acupoint administrated to achieve more efficient and safe treatment of bronchial asthma. Meanwhile, based on the hypothesis of “acupoint reservoir response”, the model of bronchial asthma in guinea pigs was established, sinapine thiocyanate was used as a model drug, microdialysis technology was used to study the pharmacokinetics and pharmacodynamics of sinapine thiocyanate, PK/PD model was established of the target site. So as to reveal the mechanism of self-dissolving microneedle, the relationship between local absorption and overall effect was analysised, and the drug release characteristics was investigated. In this study, based on the theory of an interal illness treated by external treatment, the way of the traditional clinical acupoint administration was broken through, a unique method of administration and scientific basis for Chinese medicine treatment of internal diseases was offered, the principle of traditional Chinese medicine external treatment was enriched, and a new direction for the traditional Chinese medicine modernization was greated.

创建新型穴位给药方式并探索其相应的作用机制，可为慢性哮喘的治疗提供新的治疗思路及科学依据。据此，本研究将制备载药自溶性微针，以该剂型为基础进行穴位给药实现更加高效安全的治疗。同时，以“穴位贮库应答”假说为基础，建立豚鼠支气管哮喘模型，选择芥子碱硫氰酸盐为模型药物，并采用微透析技术进行药动学及药效学研究，建立靶部位PK/PD模型，分析自溶性微针穴位给药后，局部吸收与整体效应关系，探究自溶性微针用于穴位给药的释药效应及规律，从而揭示其作用机制。本研究基于内病外治之理论，突破传统临床穴位给药的思维定式，为中药内病外治给药系统研究提供独特的给药方法及科学依据，丰富中医内病外治法的原理，为中医中药走向现代化开辟新方向。

本研究创建了一种新型穴位给药方式并探索这种新型给药系统及白芥子散对过敏性哮喘的治疗作用，可为慢性哮喘的治疗提供新的治疗思路及科学依据。主要研究内容及结果如下：（1）以芥子碱硫氢酸盐为研究对象，制备芥子碱硫氢酸盐-自溶性微针，考察其理化性质，再进行体外释放和在体透皮实验，考察药物的透皮效果进行芥子碱硫氰酸盐自溶性微针肺部药效学研究，为支气管哮喘治疗药物研究及穴位给药研究奠定药动学基础。所制备的ST-AMN微针具有足够的机械强度刺穿皮肤角质层并具有良好的缓释效果。（2）采用UPLC-MS 法测定豚鼠肺部微透析液中芥子碱硫氰酸盐含量，分别建立不同给药途径的药-时曲线、获得药动学参数、确认隔室模型。建立了卵白蛋白（OVA）诱导的豚鼠支气管哮喘模型，釆用微透析技术从支气管哮喘模型豚鼠的肺部取样，选用组胺、IL-10、TNF-α做为局部药效学指标，评价了芥子碱硫氰酸盐自溶性微针治疗支气管哮喘的药效学，药效学结果表明芥子碱硫氰酸盐自溶性微针具有显著的抑制豚鼠支气管哮喘的作用。（3）对口服给药组、穴位给药组的药动学和药效学数据分别进行分析，最终拟合出芥子碱硫氰酸盐口服、穴位给药肺部结合模型的参数，实现对实验结果的评价分析，进而得到穴位、肺部药物浓度与效应之间的定量方程，预测芥子碱硫氰酸盐口服和芥子碱硫氰酸盐自溶性微针穴位给药后局部的药物浓度和效应随时间变化的规律。口服给药、穴位给药、非穴位给药得到数据显示，穴位给药途径的Emax值、AUC值、keo值、IOD值等指标，均高于普通经皮给药和口服给药组，说明穴位给药起效更慢、药物持续释放时间较长，药效更佳平缓，总体治疗水平完全可达到甚至超过口服给药，说明药物治疗作用是协同经络穴位对人体的调节功能，相互激发和迭加而产生。（4）白芥子散穴位、非穴位贴敷治疗哮喘的作用及对施药穴位皮肤的局部作用机制研究。白芥子散穴位贴敷治疗过敏性哮喘有效，可有效回调血液（血清/血浆）中Ig-E、IL-4、IFN-γ、TNF-α等指标，穴位组优于非穴位组，非穴位组治疗作用不明显。