Abstract
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.
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