基于T细胞介导炎性机制探讨中药注射剂的类过敏反应机制

Currently the more and more severe adverse reactions have turned into the obstacles which restrict the clinical application of the traditional Chinese medicine injections, we should discover the main reason and mechanism of the adverse reactions of the traditional Chinese medicine injections, so that clinical safe and rational administration of the traditional Chinese medicine injections and the healthy development of traditional Chinese medicine will come true. For the past few years, scholars have already realized that anaphylactoid reaction were the important cause of adverse reactions of the traditional Chinese medicine injections, but the specific mechanism of anaphylactoid reaction were inadequacy identified. Our laboratory have found that Shuangling injection and Paclitaxel Injection can cause anaphylactoid reactions in ICR mice, but the same dose of SHLI and Paclitaxel injection can't cause anaphylactoid reaction in SCID mice and BALB/C nude mice, so we consider that inflammatory reaction mediated by T cells may play a important role in the adverse reactions which induced by traditional Chinese medicine injections. In this case, we will ready to apply T and B cells combined immunodeficiency disease mouse-SCID mouse as animal models and introduce the technology of T subgroup cell's separation and purification, then we will inject the separated and purified CD4+CD25+T cell into SCID mouse, so we will get the animal models of T cell's immunologic reconstitution. After that, we input reconstituted cytokine as TNF-α、IL-1、IFN-γ、IL-6 which were secreted by CD4+CD25+T cell through the tail vein of SCID mouse and the animal model of differential cytokinemia will be completed too. Finally, we will compare the different regular pattern of anaphylactoid reaction of ICR mouse, SCID mouse, T cell's immunologic reconstitution model and differential cytokinemia model that be induced by Shuangling injection and paclitaxel injection, so that to illustrate the correlation of anaphylactoid reaction induced by traditional Chinese medicine injections and inflammatory reaction mediated by T cells.From the above, we can reveal the mechanism of anaphylactoid reaction's genesis based on the inflammatory reaction mediated by T cells and provide the scientific groundwork to search the prevention and remedy of anaphylactoid reaction induced by traditional Chinese medicine injections.

近年来，学者们已经认识到中药注射剂引发的类过敏反应是其发生不良反应的重要原因，但对于其具体的机制目前研究均不十分明确。本实验室曾经发现T细胞介导的炎症反应在类过敏反应中可能起到关键作用，因此本研究拟以SCID这一T、B细胞联合免疫缺陷小鼠为动物模型，采用T细胞亚群分离纯化技术，在SCID小鼠上进行T细胞免疫重建，或采用T细胞分泌的主要细胞因子TNF-α、IL-1、IFN-γ、IL-6等分别注射入SCID小鼠造成特种高细胞因子血症，观察中药注射剂（以注射用双黄连为研究对象，以西药注射剂紫杉醇注射液为对照）在上述条件下诱导类过敏反应的规律，并与在正常免疫小鼠（ICR小鼠）上中药注射剂诱导的类过敏反应规律进行比较，，以阐明T细胞介导的炎症反应与中药注射剂类过敏反应的相关性，揭示基于T细胞介导的炎症机制诱导类过敏反应发生的机理，为寻找中药注射剂类过敏反应预防和治疗方法提供科学基础。

双黄连注射剂是临床上疗效好运用广，并且不良反应发生率也较高的中药注射剂之一，本课题前期研究已证实双黄连注射液的的不良反应以类过敏反应为主。紫杉醇注射液由于其辅料临床上也常有类过敏反应的报道。本研究在本课题组前期建立的小鼠类过敏反应模型基础上，进一步量化了类过敏反应判断标准；并借助该模型，通过对双黄连注射液（SHLI）和紫杉醇注射液（PI）分别在ICR小鼠和免疫缺陷小鼠（SCID）小鼠上诱发的类过敏反应包括病理组织的比较分析，认为T细胞的确参与了双黄连和紫杉醇注射液的类过敏反应；运用磁珠分离技术分理出CD4-、CD4+CD25-和CD4+CD25+Treg T细胞并成功移植入SCID小鼠建立了T细胞免疫重建模型，比较了SHLI和PI在T细胞移植前后的SCID小鼠类过敏反应，证明T辅助细胞和T杀伤细胞均可能具有增强SHLI和PI类过敏反应的作用，而T调节细胞可抑制SHLI和PI引起的类过敏反应；并通过其中细胞因子等检测，初步认为其中T辅助细胞的增强作用主要是通过炎性因子TNF-α和IL-6而发挥的，IL-10可能是T调节细胞发挥抑制作用的主要功能因子之一；最后通过建立不同的高细胞因子血症模型，再一次验证了IL-6和TNF-α有增强SHLI和PI类过敏反应的趋势，IL-10和/或TGF-β有一定减弱SHLI和PI类过敏反应的趋势。本研究通过具有诱发类过敏反应的中西药注射剂进行对比平行研究，初步揭示了T细胞及细胞因子介导的炎症免疫作用在注射剂导致的类过敏反应中所发挥的可能作用。