Yue Zhen, Zhou Jing-Ru, Wang Chun-Sheng, Pu Xue-Li, Parehati Guligeiya, Cao Li-Wei, Bayina Batesurong, Ye Jian-Rong
Department of Anesthesiology, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, 830054, People's Republic of China.
Department of Anesthesiology, Changji People's Hospital, Changji, 831100, People's Republic of China.
J Inflamm Res. 2025 Jul 24;18:9837-9848. doi: 10.2147/JIR.S522640. eCollection 2025.
This study aimed to evaluate the effects of cyst fluid (EgCF) on the phenotypic characteristics and degranulation activity of mast cells during echinococcosis-associated hypersensitivity reactions, using the RBL-2H3 rat basophilic leukemia cell line as an in vitro model.
The cytotoxic effects of EgCF on RBL-2H3 cells were assessed using the Cell Counting Kit-8 assay. Degranulation activity was quantified by measuring β-hexosaminidase release. Morphological alterations and cytoskeletal changes were evaluated via toluidine blue staining and phalloidin-labeled confocal laser scanning microscopy. Apoptotic cell populations were quantified using flow cytometry.
EgCF at mass concentrations ranging from 0.5 to 5 mg/mL significantly enhanced cell proliferation. Higher EgCF concentrations were associated with increased morphological deformation, as evidenced by toluidine blue staining, and with significantly elevated β-hexosaminidase release compared to the control group (p < 0.01). Flow cytometry revealed a statistically significant increase in apoptosis rates in the treated groups (p < 0.01). Within 30 minutes of EgCF exposure, phalloidin staining showed notable cytoskeletal reorganization, including cellular wrinkling and pseudopodia extension. These morphological changes were partially reversed within 45 to 60 minutes.
EgCF induces morphological transformation and promotes degranulation in mast cells, with the intensity of these responses correlating positively with EgCF concentration. These findings suggest a dose-dependent role of EgCF in modulating mast cell activity during parasitic hypersensitivity responses.
本研究旨在以RBL-2H3大鼠嗜碱性白血病细胞系作为体外模型，评估棘球蚴囊液（EgCF）对棘球蚴病相关超敏反应期间肥大细胞表型特征和脱颗粒活性的影响。
使用细胞计数试剂盒-8法评估EgCF对RBL-2H3细胞的细胞毒性作用。通过测量β-己糖胺酶释放来定量脱颗粒活性。通过甲苯胺蓝染色和鬼笔环肽标记的共聚焦激光扫描显微镜评估形态学改变和细胞骨架变化。使用流式细胞术对凋亡细胞群体进行定量。
质量浓度为0.5至5 mg/mL的EgCF显著增强细胞增殖。较高的EgCF浓度与形态变形增加有关，甲苯胺蓝染色证明了这一点，并且与对照组相比，β-己糖胺酶释放显著升高（p < 0.01）。流式细胞术显示治疗组的凋亡率有统计学意义的增加（p < 0.01）。在暴露于EgCF的30分钟内，鬼笔环肽染色显示细胞骨架有明显的重组，包括细胞起皱和伪足延伸。这些形态学变化在45至60分钟内部分逆转。
EgCF诱导肥大细胞发生形态转变并促进其脱颗粒，这些反应的强度与EgCF浓度呈正相关。这些发现表明EgCF在寄生虫超敏反应期间调节肥大细胞活性中具有剂量依赖性作用。
