[Mechanism of the RhoA/ROCK2 signaling pathway in regulating the inflammatory response and migration of Listeria monocytogenes-infected macrophages].

Objective To investigate the regulatory role of Ras homolog family member A (RhoA) in the inflammatory response and migration of RAW264.7 macrophages following Listeria monocytogenes (Lm) infection. Methods RAW264.7 cells were transfected with small interfering RNA (si-RhoA) to knock down RhoA expression, followed by Lm infection at a multiplicity of infection (MOI) of 10 for 48 hours. Phagocytosis and Transwell assays were conducted to evaluate the bacterial engulfment and migratory capacities of RAW264.7 cells, respectively. The mRNA levels of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α (TNF-α), and IL-10 were quantified using quantitative real-time PCR (qRT-PCR). The secretion of these cytokines in the cell supernatants was measured by ELISA. Western blot was performed to assess the protein expression of ROCK1 and ROCK2. Results Knockdown of RhoA reduced the migration ability of RAW264.7 cells infected with Lm, suppresses the expression of IL-1β and IL-6 at both mRNA and protein levels, and had no significant effect on TNF-α or IL-10 expression and pahagocytosis. Additionally, ROCK2 protein expression was significantly decreased, while ROCK1 protein expression remained unchanged. Conclusion Knockdown of RhoA inhibits the inflammatory response and migratory capacity of Lm-infected RAW264.7 cells by blocking the ROCK2 signaling pathway, and targeting the RhoA/ROCK2 signaling pathway may potentially constitute a novel strategy against Lm infection.