基于VIP介导的肠道免疫功能失调研究健脾益气摄血方治疗ITP效应机制

At present, the pathogenesis of ITP has not yet fully elucidated. Previous studies confirmed the beneficial effect of Si Jun Zi Decoction on ITP. Our previous clinical trials show that serum vasoactive intestinal peptide in ITP patients was significantly increased after the treatment of Jianpi Yiqi Shexue Decoction in Chinese medicine, but there is no research to explore its association with VIP. Reports from Domestic and foreign studies have shown that the incidence of ITP is derived from the immune disorder attributable to the breakout of T h1/Th2 and Th17/Treg dynamic balance. Therefore, In this study, based on our established ITP mouse model, we’d like to mainly explore VIP-mediated intestinal immune regulation, the association between VIP expression, VIP level and spleen deficiency, and Th1/Th2, Th17/Treg balance; to elucidate relevant mechanism of JianpiYiqi Shexue Decoction in Chinese medicine on immune thrombocytopenia (ITP) via VIP-induced immunoregulation in order to provide the biological evidence of “the spleen manage blood” in the theory of Chinese medicine.

目前ITP发病机制至今尚未完全阐明。国内外研究认为ITP发病免疫失耐受有关。既往研究结果证实，从脾论治、以四君子汤为基础的健脾益气摄血方，治疗ITP临床疗效确切。我们前期实验证明健脾益气摄血方干预后的患者血清血管活性肠肽（VIP）明显升高，VIP 为公认的小鼠脾虚证模型造模评价标准之一。目前尚无关于VIP与ITP发生相关性报道。该课题通过建立ITP小鼠模型，以VIP介导肠道免疫调控为主线，辅以Th1/Th2及Th17/Treg免疫调节，观察ITP发生与VIP及其受体表达水平及肠道免疫失调的相关性，阐明健脾益气摄血方的效应机制，探索脾主统血脏象理论的部分生物学内涵。

免疫失耐受与ITP发病关系密切，VIP调节免疫方面起着重要作用。课题组前期研究显示，从脾论治、以四君子汤为基础的健脾益气摄血方治疗TIP临床疗效确切。本课题在前期研究基础上，通过注射APS构建免疫性血小板减少症小鼠模型，以VIP介导肠道免疫调控为主线，辅以Th1/Th2及Th17/Treg免疫调节，采用ELISA、流式细胞术、免疫浊度检测法、免疫组化法、TUNEL法等方法，对健脾益气摄血方治疗ITP的效应机制展开深入研究。结果显示健脾益气摄血方可通过调节VIP表达水平、提升肠黏液sIgA、sIgG含量、降低肠系膜淋巴结淋巴组织P53蛋白及淋巴细胞凋亡率，改善肠道免疫功能，同时还可改善Th1/Th2、Th17/Treg免疫失衡，从而发挥治疗ITP作用。本研究基本探明ITP发生与VIP表达水平及肠道免疫失调的相关性，以及健脾益气摄血方的效应机制，为健脾益气摄血方治疗ITP提供了科学依据。