Pretreatment with R(+)-verapamil significantly reduces mortality and cytokine expression in murine model of septic shock

It is well known that cytokines play an important role in the pathogenesis of sepsis and septic shock. There is evidence indicating that the membrane transporter, P-glycoprotein (P-gp), may be involved in the release of cytokines, such as IL-2, IL-4 or IFN-γ. The aim of this study was to assess the influence of P-gp inhibitor, R(+)-verapamil, on cytokine expression in serum and tissues as well as survival rate of mice with LPS-induced septic shock. These effects were compared with the response to treatment with pentoxifylline, lisofylline, and prednisolone administered alone or after pretreatment with R(+)-verapamil. When given as a single agent, R(+)-verapamil significantly decreased serum levels of TNF-α and IFN-γ and protected mice from endotoxin lethality. Moreover, it decreased up-regulated by LPS TNF-α gene expression in the liver and lungs. Given concomitantly with immunomodulatory compounds, it enhanced their beneficial impact on the survival of mice with septic shock. The highest increase in survival rate was observed in combination with pentoxifylline (7% vs. 67%). The most striking differences observed between saline and R(+)-verapamil pretreated animals on combination therapy included down-regulation of TNF-α, higher levels of IL-6, and decreased IFN-γ concentrations. These results suggest that P-gp may be involved in the release of IFN-γ, and possibly also TNF-α, in mice with septic shock. R(+)verapamil improves survival of mice receiving a lethal dose of LPS and significantly potentiates the protective effect of pentoxifylline and prednisolone against LPS-induced lethality, probably as a result of both P-gp inhibition and a synergistic interaction at the gene level.