黄芩汤调控线粒体自噬通路逆转肠黏膜屏障损伤、Th17/Treg失衡干预aGVHD的研究

The damage of intestinal mucosa and imbalance of Th17 / Treg cells proportion after be treated with the chemotherapy regimen of the Allogeneic hematopoietic stem cell transplantation , are both the important mechanisms of acute graft versus host disease (aGVHD)as the crucial complication of Allo-HSCT.These years,there were some studies found that,oxygen free radical caused by mitochondrial autophagy imbalance can aggravate the damage of intensified intestinal mechanical barrier , and mitochondrial autophagy pathway can regulate the proportion of Th17 / Treg cells by intervening in hypoxia inducing factor.And based on article 19 pathogenesis, symptoms of aGVHD are belong to "All vomit acid, violent pouring, all belong to hot " .After we have some studies in mitochondria,this time we will establish the mouse model of aGVHD to prove the crucial rule of mitochondrial autophagy PINK1 / Parkin pathways in aGVHD,by using the in vivo and in vitro experiments,respectively.We will complish this subject in three level: tissue,cell and molecular level to. And then we will attempt to find whether huangqin soup can reduce ROS , to protect the intestinal mucosal barrier, reduce the release of inflammatory cytokines, and correction of Th17 /Treg cells imbalances,by regulating mitochondrial autophagy pathway,and eventually to interrupt the process of aGVHD.This research will offer new targets and methods for aGVHD treatment and promote the quality of life.

急性移植物抗宿主病（aGVHD）是异基因造血干细胞移植后早期致死的重要并发症之一，预处理后肠黏膜屏障损伤、Th17/Treg细胞失衡是aGVHD发病的重要机制，而线粒体自噬失衡导致的氧自由基释放失控加剧肠黏膜屏障损伤、线粒体自噬PINK1/Parkin通路可以通过干预缺氧诱导因子调控Th17/Treg细胞比例。基于病机十九条，aGVHD胃肠道症状当属于“诸呕吐酸，暴注下迫，皆属于热”的范畴。本项目结合前期对线粒体的研究成果，建立半相合造血干细胞移植aGVHD小鼠模型，通过体内、体外实验两个方面，在组织、细胞、分子三个水平，深入探讨PINK1/Parkin通路在aGVHD发病中的作用，评价黄芩汤对aGVHD的体内外药效，明确其对该通路的调控作用，阐明其逆转肠黏膜屏障损伤、Th17/Treg细胞失衡从而打断aGVHD进程的具体机制。研究将为改善aGVHD症状提供新策略，提升移植后患者生存质量。

异基因造血干细胞移植是目前根治血液系统恶性疾病的重要方法。急性移植物抗宿主病，是allo-HSCT最主要的并发症及死亡原因之一，观察到aGVHD模型小鼠小肠中存在异常的线粒体自噬，为进一步阐释aGVHD发病机制并寻找中医药干预aGVHD的具体机制，我们使用黄芩汤进行了干预，以丰富中医学“诸呕吐酸，暴注下迫，皆属于热”理论科学内涵。我们在体内、体外实验中证实了aGVHD患者异常表达的lnc-AC145676.2.1-6-3可以通过海绵体机制干预STX3，然后进一步证实其通过PTEN/AKT/mTOR通路诱导caco2细胞自噬水平增加，以上结果提示该LncRNA可作为移植后aGVHD的新型生物标志物，该部分研究为移植后肠道aGVHD的诊断、治疗提供了可靠的理论基础。然后我们构建了小鼠单倍体造血干细胞移植后aGVHD模型来研究黄芩苷、黄芩汤对肠道aGVHD的干预作用。黄芩汤及其主要有效成分黄芩苷可以通过干预STX相关通路提高LC3-II/I、Beclin1等自噬相关蛋白的表达，从而增加自噬泡的产生，促进细胞对异常物质的代谢、改善线粒体功能状态，减轻小肠黏膜上皮细胞的凋亡、脱落。经黄芩苷、黄芩汤干预后，小鼠aGVHD疾病等级明显降低，aGVHD小鼠存活时间显著延长，小肠粘膜病理评分显著下降，小肠黏膜上皮结构更加完整。阐明了黄芩汤、黄芩苷改善肠黏膜屏障的可能机制，为中药治疗肠道急性排异提供了新的选择。并在国内一级学报或核心期刊及SCI期刊上发表依托该课题的论文8篇，其中SCI 3篇，参与国际会议2次，国内会议1次，培养研究生5名。