Wang Xin, Mijiti Wubulikasimu, Jia Qiyu, Yi Zhifei, Ma Junchao, Zhou Ziyu, Xie Zengru
Department of Orthopedics and Trauma, The First Affiliated Hospital of Xinjiang Medical University, Ürümqi, Xinjiang, China.
Key Laboratory of High Incidence Disease Research in Xingjiang (Xinjiang Medical University), Ministry of Education, Ürümqi, Xinjiang, China.
Front Microbiol. 2024 Mar 27;15:1381012. doi: 10.3389/fmicb.2024.1381012. eCollection 2024.
Hydatid disease is caused by parasites and can affect various tissues and organs in the body. The disease is characterized by the presence of hydatid cysts, which contain specific antigens that interact with the host's immune system. Mesenchymal stem cells (MSCs) are pluripotent stem cells that can regulate immunity through the secretion of extracellular vesicles (EVs) containing microRNAs (miRNAs).
In this study, hydatid antigens were isolated from sheep livers and mice peritoneal cavities. MSCs derived from mouse bone marrow were treated with different hydatid antigens, and EVs were isolated and characterized from the conditioned medium of MSCs. Small RNA library construction, miRNA target prediction, and differential expression analysis were conducted to identify differentially expressed miRNAs. Functional enrichment and network construction were performed to explore the biological functions of the target genes. Real-time PCR and Western blotting were used for miRNA and gene expression verification, while ELISA assays quantified TNF, IL-1, IL-6, IL-4, and IL-10 levels in cell supernatants.
The study successfully isolated hydatid antigens and characterized MSC-derived EVs, demonstrating the impact of antigen concentration on MSC viability. Key differentially expressed miRNAs, such as miR-146a and miR-9-5p, were identified, with functional analyses revealing significant pathways like Endocytosis and MAPK signaling associated with these miRNAs' target genes. The miRNA-HUB gene regulatory network identified crucial miRNAs and HUB genes, such as Traf1 and Tnf, indicating roles in immune modulation and osteogenic differentiation. Protein-protein interaction (PPI) network analysis highlighted central HUB genes like Akt1 and Bcl2. ALP activity assays confirmed the influence of antigens on osteogenic differentiation, with reduced ALP activity observed. Expression analysis validated altered miRNA and chemokine expression post-antigen stimulation, with ELISA analysis showing a significant reduction in CXCL1 expression in response to antigen exposure.
This study provides insights into the role of MSC-derived EVs in regulating parasite immunity. The findings suggest that hydatid antigens can modulate the expression of miRNAs in MSC-derived EVs, leading to changes in chemokine expression and osteogenic capacity. These findings contribute to a better understanding of the immunomodulatory mechanisms involved in hydatid disease and provide potential therapeutic targets for the development of new treatment strategies.
包虫病由寄生虫引起，可影响身体的各种组织和器官。该疾病的特征是存在包虫囊肿，其中含有与宿主免疫系统相互作用的特定抗原。间充质干细胞（MSCs）是多能干细胞，可通过分泌含有微小RNA（miRNAs）的细胞外囊泡（EVs）来调节免疫。
在本研究中，从绵羊肝脏和小鼠腹腔中分离出包虫抗原。用不同的包虫抗原处理从小鼠骨髓中获得的间充质干细胞，并从间充质干细胞的条件培养基中分离和鉴定细胞外囊泡。进行小RNA文库构建、miRNA靶标预测和差异表达分析，以鉴定差异表达的miRNAs。进行功能富集和网络构建，以探索靶基因的生物学功能。使用实时PCR和蛋白质印迹法验证miRNA和基因表达，而ELISA测定法定量细胞上清液中TNF、IL-1、IL-6、IL-4和IL-10的水平。
该研究成功分离出包虫抗原并鉴定了间充质干细胞来源的细胞外囊泡，证明了抗原浓度对间充质干细胞活力的影响。鉴定出关键的差异表达miRNAs，如miR-146a和miR-9-5p，功能分析揭示了与这些miRNAs靶基因相关的重要途径，如内吞作用和MAPK信号传导。miRNA-HUB基因调控网络鉴定出关键的miRNAs和HUB基因，如Traf1和Tnf，表明它们在免疫调节和成骨分化中发挥作用。蛋白质-蛋白质相互作用（PPI）网络分析突出了Akt1和Bcl2等核心HUB基因。碱性磷酸酶（ALP）活性测定证实了抗原对成骨分化的影响，观察到ALP活性降低。表达分析验证了抗原刺激后miRNA和趋化因子表达的改变，ELISA分析显示，暴露于抗原后CXCL1表达显著降低。
本研究深入探讨了间充质干细胞来源的细胞外囊泡在调节寄生虫免疫中的作用。研究结果表明，包虫抗原可调节间充质干细胞来源的细胞外囊泡中miRNAs的表达，导致趋化因子表达和成骨能力的变化。这些发现有助于更好地理解包虫病所涉及的免疫调节机制，并为开发新的治疗策略提供潜在的治疗靶点。
