A Cystatin Derived from Fasciola gigantica Suppresses Macrophage-mediated Inflammatory Responses
Keywords:Cystatin, Fasciola gigantica, Inflammation, Pro-inflammatory cytokines
Introduction: The liver fluke, Fasciola gigantica, can modulate host immune responses through immunomodulatory molecule to retain a suitable environment for its long-term survival. These molecules, such as, cystatin, the cysteine cathepsin inhibitor, etc., may be used for the treatment of human inflammatory diseases.
Objectives: This study aimed to identify the potential of recombinant cystatin from Fasciola gigantica (rFgCyst) whether it can inhibit inflammation mediated by LPS-treated macrophages.
Methods: Mouse macrophage, RAW264.7 cell, and human macrophage, THP-1 cell, were cultured in 96-well plates with various concentration of rFgCyst. Cell viability was determined by MTT assay. For anti-inflammatory assay, RAW264.7 cells and THP-1 cells were treated with rFgCyst for 24 h, followed by inflammation stimulating with LPS (E. coli 0111: B4) and additionally incubating for 12 and 24 h, respectively. After incubation, the culture media and the cells were collected to monitor the levels of pro-inflammatory cytokine including IL6, IL1β, and TNF-α and its mRNAexpression using ELISA, western blot and quantitative real-time RT-PCR (qRT-PCR), respectively.
Results: Pretreatment of RAW264.7 cells and THP-1 cell with rFgCyst inhibited the production of pro‑inflammatory cytokines including IL6, IL1β, and TNF-α induced by LPS both gene and protein expression levels in dose- and time-dependent manner.
Conclusions: rFgCyst reduces inflammation in in vitro model probably by reducing pro-inflammatory cytokines. These results suggest that rFgCyst may have a potential function against human inflammatory diseases in future.
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