Song Jialin, Han Yanxiao, Chen Yuyan, Cheng Lin, Xiao Juan, Li Ai, Kong Dexiao, Jiang Yang, Zheng Chengyun
Department of Hematology, The Second Hospital of Shandong University, Jinan, Shandong, China.
Institute of Biotherapy for Hematological Malignancy, Shandong University, Jinan, Shandong, China.
Stem Cell Res Ther. 2025 Jul 1;16(1):337. doi: 10.1186/s13287-025-04443-x.
Hemorrhagic cystitis (HC), a frequent complication of hematopoietic stem cell transplantation (HSCT), significantly affects quality of life and may worsen prognosis. Mesenchymal stem cells (MSCs) are known for their anti-inflammatory and tissue-regenerative properties. IL-1 receptor antagonist (IL-1Ra) blocks IL-1α and IL-1β by binding IL-1 receptors, offering potential therapeutic benefits. The aim of this study was to explore the therapeutic effect of MSCs overexpressing IL-1Ra on HC and investigate the underlying mechanisms.
MSCs were isolated from human umbilical cord tissues, and IL-1Ra-overexpressing MSCs (oeIL-1Ra-MSCs) were generated using lentiviral transfections. HC was induced in rats by cyclophosphamide administration. Rats received tail vein injections of either oeIL-1Ra-MSCs or control MSCs (Mock-MSCs). Hematuria and bladder tissue changes were assessed using test strips and hematoxylin & eosin (HE) staining. Immunohistochemistry detected molecular changes in bladder tissues. Gene expression differences between the two MSC groups were analyzed by mRNA sequencing and ChIP techniques.
Treatment with oeIL-1Ra-MSCs significantly alleviated hematuria and reduced bladder edema and hemorrhage, and reduced mRNA expression levels of IL-1β, IL-6, and TNF-α in bladder tissues, compared with those in the Mock-MSC treatment group. Immunohistochemical staining showed a higher presence of CD105-positive cells (a marker for human MSCs) and CD31-positive vessels in bladder tissues treated with oeIL-1Ra-MSCs, indicating enhanced MSC migration and vascular stability. In vitro migration assay demonstrated a higher migration capacity of IL-1Ra overexpressing MSCs compared with that of control MSCs. Moreover, angiopoietin-1 (Ang-1) expression increased, while Angiopoietin-2 (Ang-2) expression decreased in bladder tissues treated with oeIL-1Ra-MSCs, suggesting enhanced blood vessel stabilization. Conditioned medium from oeIL-1Ra-MSC cultures stimulated human umbilical vein endothelial cell (hUVEC) migration, proliferation, and angiogenesis more effectively compared with that in control MSCs. mRNA sequencing revealed elevated HtrA3 expression in oeIL-1Ra-MSCs compared with that in control MSCs. Molecular analysis suggested that IL-1Ra overexpression in MSCs upregulated HtrA3 expression through inhibition of the JNK-c-Jun pathway and activation of the ERK-Egr-1 pathway.
Overexpression of IL-1Ra significantly enhances the therapeutic efficacy of MSCs in HC by promoting MSC migration to damaged bladder tissues, suppressing inflammation, stabilizing blood vessels, and upregulating angiogenesis via activation of HtrA3 signaling pathways.
出血性膀胱炎（HC）是造血干细胞移植（HSCT）常见的并发症，严重影响生活质量并可能使预后恶化。间充质干细胞（MSCs）以其抗炎和组织再生特性而闻名。白细胞介素-1受体拮抗剂（IL-1Ra）通过结合IL-1受体来阻断IL-1α和IL-1β，具有潜在的治疗益处。本研究旨在探讨过表达IL-1Ra的MSCs对HC的治疗效果并研究其潜在机制。
从人脐带组织中分离出MSCs，并通过慢病毒转染生成过表达IL-1Ra的MSCs（oeIL-1Ra-MSCs）。通过给予环磷酰胺诱导大鼠发生HC。大鼠经尾静脉注射oeIL-1Ra-MSCs或对照MSCs（Mock-MSCs）。使用试纸条和苏木精-伊红（HE）染色评估血尿和膀胱组织变化。免疫组织化学检测膀胱组织中的分子变化。通过mRNA测序和染色质免疫沉淀技术分析两组MSCs之间的基因表达差异。
与Mock-MSC治疗组相比，oeIL-1Ra-MSCs治疗显著减轻了血尿，减少了膀胱水肿和出血，并降低了膀胱组织中IL-1β、IL-6和TNF-α的mRNA表达水平。免疫组织化学染色显示，在oeIL-1Ra-MSCs处理的膀胱组织中，CD105阳性细胞（人MSCs的标志物）和CD31阳性血管的数量更多，表明MSCs迁移增强和血管稳定性提高。体外迁移试验表明，过表达IL-1Ra的MSCs比对照MSCs具有更高的迁移能力。此外，在oeIL-1Ra-MSCs处理的膀胱组织中，血管生成素-1（Ang-1）表达增加，而血管生成素-2（Ang-2）表达减少，表明血管稳定性增强。与对照MSCs相比，oeIL-1Ra-MSC培养物的条件培养基更有效地刺激人脐静脉内皮细胞（hUVEC）的迁移、增殖和血管生成。mRNA测序显示，与对照MSCs相比，oeIL-1Ra-MSCs中HtrA3表达升高。分子分析表明，MSCs中IL-1Ra的过表达通过抑制JNK-c-Jun途径和激活ERK-Egr-1途径上调了HtrA3表达。
IL-1Ra的过表达通过促进MSCs向受损膀胱组织迁移、抑制炎症、稳定血管以及通过激活HtrA3信号通路上调血管生成，显著增强了MSCs对HC的治疗效果。
