Qiu Zhiyuan, Liu Hui, Cao Rongxuan, Wang Shan, Wang Junjun, Xu Wenjun, Zhang Rui, Wang Baohong, Zhang Xiaoting, Li Qianpeng
Department of Hematology, Weifang People's Hospital, Shandong Second Medical University, Weifang, China.
Department of Cardiology, Weifang People's Hospital, Shandong Second Medical University, Weifang, China.
Immunology. 2025 Aug;175(4):534-543. doi: 10.1111/imm.13935. Epub 2025 May 22.
Haematopoietic stem cell transplantation (HSCT) is one of the key strategies for treating various haematologic malignancies. Although there are haematopoietic stem cells (HSCs) in umbilical cord blood (UCB), the number is limited. Thus, the purpose of this work was to investigate if endogenous hydrogen sulphide (HS) could encourage the ex vivo expansion of HSCs produced from UCB (UCB-HSCs). The CD34 and CD34CD38 cells were enriched and separated by immunomagnetic beads. UCB-HSCs were treated with overexpression plasmids of β-synthase (CBS), cystathionine γ-lyase (CSE), 3-mercaptopyruvate sulphurtransferase (MPST) and/or stimulated with AG490 (JAK2/STAT3 inhibitor) for 4, 7 or 10 days, respectively. The content of HS in cells was detected using its assay kit. The proportion and quantity of CD34, CD34CD38 and CXCR4CD34 cells as well as the ALDH1A1CD34 cells in CD34 cells were detected by flow cytometry. qPCR was used to detect the expression of CD34, CXCR4 and ALDH1A1 in CD34 cells. Western blot was used to detect the activation of the JAK2/STAT3 pathway in CD34 cells. The results showed that endogenous HS enhanced the ex vivo expansion of CD34 and CD34CD38 cells, upregulated the expression of CXCR4 and ALDH1A1 during ex vivo expansion of HSCs, and promoted the JAK2/STAT3 pathway in CD34 cells. However, the aforementioned effects of endogenous HS were partially reversed by AG490. In conclusion, endogenous HS promotes the activation of the JAK2/STAT3 pathway to facilitate the ex vivo expansion of UCB-HSCs.
造血干细胞移植（HSCT）是治疗各种血液系统恶性肿瘤的关键策略之一。尽管脐带血（UCB）中存在造血干细胞（HSC），但其数量有限。因此，本研究的目的是探讨内源性硫化氢（HS）是否能促进UCB产生的HSC（UCB-HSC）的体外扩增。通过免疫磁珠富集并分离CD34和CD34CD38细胞。分别用β-合酶（CBS）、胱硫醚γ-裂解酶（CSE）、3-巯基丙酮酸硫转移酶（MPST）的过表达质粒处理UCB-HSC和/或用AG490（JAK2/STAT3抑制剂）刺激4、7或10天。使用检测试剂盒检测细胞中HS的含量。通过流式细胞术检测CD34细胞中CD34、CD34CD38和CXCR4CD34细胞以及ALDH1A1CD34细胞的比例和数量。采用qPCR检测CD34细胞中CD34、CXCR4和ALDH1A1的表达。采用蛋白质免疫印迹法检测CD34细胞中JAK2/STAT3信号通路的激活情况。结果表明，内源性HS增强了CD34和CD34CD38细胞的体外扩增，在HSC体外扩增过程中上调了CXCR4和ALDH1A1的表达，并促进了CD34细胞中JAK2/STAT3信号通路。然而，AG490部分逆转了内源性HS的上述作用。总之，内源性HS通过促进JAK2/STAT3信号通路的激活来促进UCB-HSC的体外扩增。
