State Key Laboratory of Component-based Chinese Medicine, Tianjin University of Traditional Chinese Medicine, No. 10 Poyanghu Road, West Area, Tuanbo New Town, Jinghai District, Tianjin 301617, China; Research and Development Center of TCM, Tianjin International Joint Academy of Biotechnology & Medicine, 220 Dongting Road, TEDA, Tianjin 300457, China.
State Key Laboratory of Experimental Hematology, Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China.
Biomed Pharmacother. 2022 Apr;148:112675. doi: 10.1016/j.biopha.2022.112675. Epub 2022 Feb 17.
Acute graft-versus-host disease (aGVHD) is a major life-threatening complication after Allogeneic Hematopoietic Stem Cell Transplant (allo-HSCT). Although a series of immunosuppressant agents are routinely used as the first-line prevention, the morbidity and mortality rate remains high in allo-HSCT recipients. Our previous work indicated that combining Xuebijing (XBJ) with Cyclosporin A (CSA) is superior to CSA alone in preventing aGVHD. However, it was not clear which compounds in XBJ may prevent aGVHD. Whether the effective compounds in XBJ can be safely combined with CSA to prevent GVHD remain to be evaluated. Here, we accessed whether the combination of four main components in XBJ (C0127) had the same efficacy as XBJ in preventing aGVHD. In addition, the effectiveness of a novel combination therapy (C0127 + CSA) on aGVHD prophylaxis was evaluated using 16 s rRNA sequencing and RNA sequencing approaches in vitro and in vivo. In aGVHD mice, C0127 enhanced the preventive effects of CSA including decreasing mortality, maintaining weight, reducing GVHD score and reducing the expression of IL-6 and TNF-α in serum. Fatal GVHD is a frequent consequence of intestinal tract damage. We found combining C0127 with CSA alleviated the gut damage and maintained the normal physiological function of intestine by H&E staining, intestinal permeability and short chain fatty acid (SCFA) assays. Next, 16 S sequencing analysis of feces showed the combination treatment maintained the intestinal microbial diversity, normalized the intestinal microorganism and prevented flora disorder by reducing the relative abundances of Escherichia coli and Enterococcus. Further, RNA-seq analysis of colonic epithelium revealed C0127 combined with CSA chiefly regulated chemokines and cytokines in IL-17 signaling pathway. The combination treatment reduced the expression of G-CSF and its effector STAT3 (an axis that aggravated gut inflammation and flora disorder) in gut epithelium on mRNA and protein level. These findings indicated that C0127 improved the prevention of CSA in aGVHD mice partially by protecting the gut from damage through normalizing G-CSF signaling, which regulates the intestinal microbiota and the integrity of the epithelial barrier.
急性移植物抗宿主病（aGVHD）是异基因造血干细胞移植（allo-HSCT）后的一种主要危及生命的并发症。尽管一系列免疫抑制剂被常规用于一线预防，但 allo-HSCT 受者的发病率和死亡率仍然很高。我们之前的工作表明，将血必净（XBJ）与环孢素 A（CSA）联合使用在预防 aGVHD 方面优于 CSA 单独使用。然而，尚不清楚 XBJ 中的哪些化合物可以预防 aGVHD。XBJ 中的有效化合物是否可以与 CSA 安全联合用于预防 GVHD 仍有待评估。在这里，我们评估了 XBJ 的四种主要成分（C0127）的组合是否与 XBJ 具有相同的预防 aGVHD 的疗效。此外，我们还通过 16s rRNA 测序和 RNA 测序方法，在体外和体内评估了新型联合治疗（C0127+CSA）预防 aGVHD 的效果。在 aGVHD 小鼠中，C0127 增强了 CSA 的预防作用，包括降低死亡率、维持体重、降低 GVHD 评分以及降低血清中 IL-6 和 TNF-α的表达。致命性 GVHD 是肠道损伤的常见后果。我们发现，通过 H&E 染色、肠道通透性和短链脂肪酸（SCFA）测定，将 C0127 与 CSA 联合使用可以减轻肠道损伤并维持肠道的正常生理功能。接下来，粪便 16S 测序分析表明，联合治疗通过降低大肠杆菌和肠球菌的相对丰度，维持肠道微生物多样性，使肠道微生物正常化，并预防菌群失调。此外，结肠上皮的 RNA-seq 分析表明，C0127 联合 CSA 主要通过调节 IL-17 信号通路中的趋化因子和细胞因子来调节肠道微生物群。联合治疗降低了 G-CSF 及其效应物 STAT3（加重肠道炎症和菌群失调的轴）在肠道上皮的 mRNA 和蛋白水平上的表达。这些发现表明，C0127 通过保护肠道免受损伤，部分改善了 CSA 在 aGVHD 小鼠中的预防作用，通过使 G-CSF 信号正常化来调节肠道微生物群和上皮屏障的完整性。
