旋毛虫成虫体抗原对人血管内皮细胞血管生成、细胞增殖、迁移和假毛细血管形成的影响。
Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells.
机构信息
Zoonotic Diseases and One Health Group, IBSAL-CIETUS (Biomedical Research Institute of Salamanca Research Centre for Tropical Diseases), Faculty of Pharmacy, University of Salamanca, 37007, Salamanca, Spain.
Department of Physiology and Pharmacology, Biomedical Research Institute of Salamanca (IBSAL), University of Salamanca, Salamanca, Spain.
出版信息
Parasit Vectors. 2023 Mar 16;16(1):105. doi: 10.1186/s13071-023-05726-z.
BACKGROUND
Angiogenesis is defined as the formation of new vessels by sprouting of endothelial cells from pre-existing vessels in response to stimuli, such as hypoxia or inflammation. Subcutaneous dirofilariasis, caused by Dirofilaria repens, is a zoonotic disease characterized by the formation of subcutaneous nodules with the presence of at least one encapsulated worm, showing perivascular vascularization around it. The aim of this study is to analyze whether the somatic antigen of adult D. repens worms interacts with and modulates the angiogenic mechanism, cell proliferation and migration, and formation of pseudo-capillaries.
METHODS
The expression of VEGF-A, VEGFR-1/sFlt, VEGFR-2, mEnd and sEnd in cultures of human vascular endothelial cells stimulated with somatic antigen of adult worms of D. repens (DrSA), vascular endothelial growth factor (VEGF) and DrSA + VEGF were evaluated by using ELISA commercial kits. Cellular viability was analyzed by live cell count, cytotoxicity assays by using a commercial kit, cell proliferation by MTT-based assay, cell migration by wound-healing assay carried out by scratching wounds and capacity of formation of pseudo-capillaries analyzing cell connections and cell groups in Matrigel cell cultures. In all cases unstimulated cultures were used as controls.
RESULTS
DrSA + VEGF significantly increased the expression of VEGF-A, VEGFR-2 and mEndoglin compared to other groups and unstimulated cultures. Moreover, DrSA + VEGF produced cell proliferation and migration and increased the formation of pseudo-capillaries.
CONCLUSIONS
Somatic antigen of adult D. repens worms activated the proangiogenic mechanism, cell proliferation and cell migration as well as formation of pseudo-capillaries in this in vitro human endothelial cell model. These processes could be related to the survival of adult D. repens in subcutaneous nodules in infected hosts.
背景
血管生成是指内皮细胞从预先存在的血管中发芽形成新的血管，以响应缺氧或炎症等刺激。由犬恶丝虫引起的皮下丝虫病是一种人畜共患病，其特征是形成皮下结节，至少有一条包裹的虫体，其周围有血管周围血管化。本研究旨在分析成年犬恶丝虫的体抗原是否与调节血管生成机制、细胞增殖和迁移以及假毛细血管形成相互作用。
方法
使用 ELISA 试剂盒评估刺激人血管内皮细胞培养物的成年犬恶丝虫体抗原（DrSA）、血管内皮生长因子（VEGF）和 DrSA+VEGF 对 VEGF-A、VEGFR-1/sFlt、VEGFR-2、mEnd 和 sEnd 的表达。通过活细胞计数分析细胞活力，通过使用商业试剂盒进行细胞毒性测定，通过 MTT 基于测定法进行细胞增殖测定，通过划痕伤口进行细胞迁移测定，通过分析 Matrigel 细胞培养物中的细胞连接和细胞群来分析假毛细血管形成的能力。在所有情况下，未刺激的培养物均用作对照。
结果
与其他组和未刺激的培养物相比，DrSA+VEGF 显著增加了 VEGF-A、VEGFR-2 和 mEndoglin 的表达。此外，DrSA+VEGF 产生了细胞增殖和迁移，并增加了假毛细血管的形成。
结论
成年犬恶丝虫的体抗原激活了该体外人内皮细胞模型中的促血管生成机制、细胞增殖和细胞迁移以及假毛细血管形成。这些过程可能与感染宿主皮下结节中成年犬恶丝虫的存活有关。
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