Alveolar echinococcosis (AE) caused by Echinococcus multilocularis (E. multilocularis), characterized by lesions composed of an aggregate of microcysts embedded in a granulomatous host's reaction. The periphery of parasite granulomas often additionally displays fibrotic reactions of varying intensity, in which E. multilocularis microenvironment fibroblasts (EMFs) laid down collagen. However, the regulation of EMFs by the infiltration of E. multilocularis microcyst fluid (MF) into granulomas remains poorly defined. This study aimed to investigate the effect of MF on migration and invasion of primary isolated EMFs cells. A mouse model of secondary infection with AE was established, and the model construction was evaluated by HE staining. EMFs were cultured in primary by tissue block adherency method. The isolated cells were identified by qPCR, immunofluorescence and Western blot. Then CCK-8 assay, cell migration/invasion assay and flow cytometry were performed to detect the effects of MF on the proliferation, migration, invasion and cell cycle of EMFs, respectively. The expressions of MMP2 and MMP9 at mRNA and protein levels in EMFs were detected by RT-qPCR and Western blot. The effect of PI3K-Akt signal transduction pathway on regulating the expression of MMPs expression was assessed by Western blot. As indicated from the results, EMFs were successfully isolated from the E. multilocularis microenvironment and identified as myofibroblasts. MF significantly facilitated the proliferation and cell cycle progression of EMFs. In addition, MF significantly improved the migration and invasion of EMFs. MF was further confirmed to up-regulate mRNA and protein expressions of MMP2 and MMP9 in EMFs, which was related to the activation of the PI3K-Akt signaling pathway. The present study demonstrates that MF can promote the migration and invasion of EMFs cells significantly, which might be via activating PI3K-Akt signaling pathway.

由多房棘球绦虫( E. multilocularis ) 引起的肺泡棘球蚴病 (AE) ，其特征是由嵌入肉芽肿宿主反应中的微囊聚集体组成的病变。寄生虫肉芽肿的外围通常还表现出不同强度的纤维化反应，其中多房大肠杆菌微环境成纤维细胞 (EMF) 沉积胶原蛋白。然而，多房大肠杆菌的渗透对 EMF 的调节微囊液 (MF) 进入肉芽肿仍然不明确。本研究旨在研究MF对原代分离的EMFs细胞迁移和侵袭的影响。建立AE继发感染小鼠模型，并通过HE染色评价模型构建。通过组织块粘附法在原代培养 EMF。通过qPCR、免疫荧光和蛋白质印迹鉴定分离的细胞。然后通过CCK-8实验、细胞迁移/侵袭实验和流式细胞术分别检测MF对EMFs增殖、迁移、侵袭和细胞周期的影响。通过RT-qPCR和Western blot检测EMFs中MMP2和MMP9在mRNA和蛋白水平的表达。通过蛋白质印迹评估PI3K-Akt信号转导通路对调节MMPs表达的影响。结果表明，EMFs 成功地从E. 多房微环境并鉴定为肌成纤维细胞。MF显着促进了EMF的增殖和细胞周期进程。此外，MF显着改善了EMF的迁移和入侵。进一步证实MF上调EMFs中MMP2和MMP9的mRNA和蛋白表达，这与PI3K-Akt信号通路的激活有关。本研究表明MF可以显着促进EMFs细胞的迁移和侵袭，这可能是通过激活PI3K-Akt信号通路。