变应性鼻炎体质模型的骨髓干细胞动员归巢及中药干预机制

Stem cells in the bone marrow (mainly MSCs and EPCs) mobilized into the peripheral circulation and directional migration / to the lesion plays a key role in the injury repair process in the diseased tissue in the homing. TCM believes that "kidney main bone marrow", deficiency of kidney yang and bone marrow microenvironment changes closely related, kidney tonifying herbs can for the treatment of allergic rhinitis has already been confirmed by clinical and experimental. But the traditional Chinese medicine to intervene the relationship between bone marrow stem cells mobilization and homing of allergic rhinitis and the occurrence and development is not clear. Our previous experimental work found that bone marrow IL-3, IL-5, EOT and other cold sex constitution model of allergic rhinitis in cytokine easier to collect, chemotaxis and activation of EOS, and the compound Chinese medicine for reinforcing kidney can intervene the change mechanism of good. According to this hypothesis, traditional Chinese medicine compound transfer / to the nasal mucosa orientation in bone marrow stem cells mobilization and homing into peripheral circulation play a key role in the process of tissue repair of nasal mucosa. We will through the cold and hot constitution model of allergic rhinitis in traditional Chinese medicine, before and after the intervention, through the detection of cell cycle and apoptosis by flow cytometry, stem cell isolation and culture and fluorescence labeling technology, study on the mechanism of allergic rhinitis by different physical models of bone marrow stem cell mobilization homing, exploring the mechanism of physical fitness and bone marrow stem cells the relationship between the mobilization and homing of traditional Chinese medicine intervention.

骨髓干细胞（主要是MSCs与EPCs）动员进入外周循环并且向病变部位定向迁移/归巢在病变组织的损伤修复过程中起关键作用。中医认为“肾主骨生髓”，肾阳虚与骨髓微环境变化密切相关，补肾中药能治疗变应性鼻炎早已被临床和实验所证实。但中药如何干预骨髓干细胞的动员归巢和变应性鼻炎发生发展的关系还不明确。我们前期实验发现变应性鼻炎的寒性体质模型中骨髓IL-3、IL-5、EOT等细胞因子更容易募集、趋化和激活EOS，而补肾中药能够很好的干预这种变化机制。据此提出假设，中药复方在骨髓干细胞动员进入外周循环并且向鼻腔黏膜定向迁移/归巢在鼻黏膜组织修复的过程中起关键作用。我们将通过建立变应性鼻炎的寒热体质模型，在中药干预前后，通过流式细胞术检测细胞周期与凋亡，干细胞分离与培养以及荧光标记等技术，进行不同体质模型的变应性鼻炎骨髓干细胞动员归巢的机制研究，探索体质与骨髓干细胞动员归巢之间的关系以及中药干预的机制。

目的：研究不同体质的变应性鼻炎的骨髓干细胞（MSCs、EPCs）的增殖、动员和归巢的规律，并探讨不同体质下中药干预的机制。.方法：通过“自然筛选－药物强化－同种体质交配传代”的方法构建SD大鼠的寒、热、常体的遗传性体质模型，在此基础上构建变应性鼻炎的疾病模型。每种体质模型分干预和非干预组，非干预组不做干预处理，常体组和寒体组均用附子理中汤干预，热体组用黄连解毒汤干预，在不同的时间节点（0、1、3、7天）取材，每个时间节点每组大鼠6只，取骨髓和血液，流式细胞检测骨髓和循环系统中干细胞（MSCs、EPCs）的含量。另取常体无干预大鼠6只，作为荧光标记 MSCs、EPCs 供体进行干细胞培养，经培养扩增后的MSCs、EPCs以荧光染料BrdU标记后，在每组各6只完成体质鼻炎造模后即经尾静脉注入循环系统。在治疗后第7天取鼻腔黏膜组织，免疫荧光检测鼻腔黏膜经BrdU标记的干细胞的含量。.结果：1.成功构建了体质的遗传模型和变应性鼻炎动物模型；2寒体组的症状比常体、热体组都严重（P＜0.05）且自然状态下骨髓干细胞的含量最少；3.寒、热、常体的中药治疗均有效果；4.三种体质不干预的情况下，各组的骨髓干细胞随着时间的推移会逐渐减少；各组的血液干细胞的动员也会随着时间的推移逐渐减少；各组的干细胞在鼻腔的归巢无影响（P＞0.05）；5.中药干预能促进各体质大鼠骨髓干细胞在骨髓中的增殖和向循环系统的动员。各体质干预后组间的归巢差异不明显（P＞0.05），但寒、常体的干预能促进干细胞向鼻腔黏膜的归巢（P＜0.05）。热体组的干预能提高EPCs的归巢（P＜0.05），但不能提高MSCs归巢（P＞0.05）。.结论：体质决定了变应性鼻炎的骨髓干细胞的表达，中药干预的机制是促进骨髓干细胞在骨髓中的增殖和向循环系统动员以及向鼻腔黏膜的归巢。