血必净在异基因造血干细胞移植后防治急性GVHD的免疫调节作用及其对受者肠道菌群的影响

Gut injury caused by chemotherapy and/or radiation is a major cause of acute Graft versus host disease (aGvHD) after Allogeneic hematopoietic stem cell transplantation (AHSCT).Current aGvHD prevention strategy targeting T cells showed limited clinical success and can cause adverse effects and toxicity. Combination of Chinese and Western medicine is promising to prevent severe aGvHD through systematic intervention that not only prevents T cell activation, controls inflammation but also fortifies the.bodies of recipients. Xuebijing (XBJ) is an effective Chinese medicine injection to treat both Sepsis and Septic Shock. Our preliminary studies revealed that Xuebijing not only promotes regulatory T cell differentiation but also prevents Th17 cell differentiation and blood clotting. In addition, it improves viability of murine macrophages while inhibits NF-kB activation. IPA signaling network analysis predicted that major ingredients of XBJ target key aGVHD signaling pathways. Based on current evidence, we hypothesize that XBJ prevents aGVHD through containing inflammation and protecting intestine to maintain gut microbiota. We choose in vitro cell based model and a murine aGVHD model to test our hypothesis. We will also determine the effect of XBJ in combination with Cyclosporin A. We will utilize cutting-edge technologies such as cytokine array, RNAseq and Flow cytometer analysis to determine the effect of XBJ on the expression of pro-inflammatory cytokines, activation of dendritic cells and T cells, and gut microbiota. With respect to expected outcomes, this project is going to have positive impact on the application of Xuebijing in the field of allogenic hematopoietic transplantation as well as the field of organ transplantation and immunotherapy.

放化疗预处理造成的肠道损伤是恶化急性移植物抗宿主病(GVHD)的重要原因，单靶点化疗方案难以预防GVHD，并产生一定的毒副作用。因此，中西医结合多靶点系统防治的策略更具解决这一难题的潜力。中药血必净具有活血化瘀，抑炎，免疫调节，拮抗内毒素，器官保护等一系列系统防治GVHD的特质。前期研究表明血必净不仅增强细胞活力，抑制过度免疫激活，而且调控T细胞分化。其组分可调控GVHD关键靶蛋白和信号通路。因此，我们提出血必净通过降低炎症反应，保护肠道组织，维持肠道菌群平衡系统防治急性GVHD的假说。本项目拟选用细胞和小鼠急性GVHD模型检验这一假说，探索血必净和环孢素A联合防治GVHD的新途径。我们将使用炎性因子阵列，流式分析，RNA测序，网络药理等方法检测血必净对炎性因子表达，免疫激活，和肠道微环境的影响，阐明多靶点系统防治GVHD的科学性，为血必净在器官移植和免疫治疗领域的应用铺垫道路。

放化疗预处理造成的肠道损伤是恶化急性移植物抗宿主病(aGVHD)的重要原因，单靶点化疗方案难以预防GVHD，并可能产生毒副作用。多靶点系统防治GVHD的策略更具潜力。中药血必净具有活血化瘀，抑炎，免疫调节，拮抗内毒素，器官保护等系统防治GVHD的特质。初步研究表明血必净增强细胞活力，抑制过度免疫激活，调控T细胞分化。其组分可调控GVHD关键靶蛋白和信号通路。因此，我们提出血必净通过降低炎症反应，保护肠道组织，维持肠道菌群平衡，系统防治急性肠道GVHD的假说。本项目选用细胞模型和小鼠急性GVHD模型检验这一假说，探索血必净和环孢素A联合防治GVHD的新途径。使用炎性因子阵列，流式分析，RNA测序，网络药理等方法检测血必净对炎性因子表达，免疫激活，和肠道微环境的影响。我们的网络药理研究表明，血必净涵盖比阳性对照药丹参注射液更多的GVHD防治靶点，包括与环孢素A共同的GVHD靶点。体外实验表明血必净可以促进血管新生，提高小鼠巨噬细胞的活力，并且有效抑制血小板聚集和黏附。此外，血必净可以与环孢素安全联用，有效提高aGVHD小鼠的生存，对异基因造血干细胞植入没有不良影响。我们的近期结果显示，血必净和低剂量环孢素联用比环孢素单独用药更有效地提高aGVHD小鼠的生存，在纲、属和种等不同水平保护肠道菌群的多样性，维持肠道菌群丰度，保护肠道组织，降低炎症因子风暴，我们还发现血必净干预有效抑制大肠杆菌和肠球菌等肠道GVHD生物标志物的相对丰度。以上结果表明，网络药理和实验药理相结合的方法可用于开发新的中西医结合疾病防治方案，保护肠道微环境有益于防治急性肠道移植物抗宿主病，中西医结合可能成为防治移植物抗宿主病的新策略。血必净和低剂量环孢素联用可系统性地防治急性移植物抗宿主病，可望提高异基因造血干细胞受者的生活质量，此研究为血必净在器官移植领域的应用铺垫了道路。