The Anti-inflammatory Effects And Mechanisms Of Salvia Miltiorrhiza Lipid-soluble Extract In Vitro And In Vivo

Aim of the study:Salvia miltiorrhiza, a traditional Chinese herbal medicine, used to treat various inflammatory diseases. In the present study, we have demonstrated the anti-inflammatory effects of S. miltiorrhiza lipid-soluble extract (SMLE) in LPS-treated RAW 264.7 cells and in endotoxemia, sepsis and PMA-induced ear edema mice along with its underlying mechanism of action.Materials and methods:RAW 264.7 cells were pretreated with vehicle or SMLE and then exposed to lipopolysaccharide (LPS) for the time indicated. Nitric oxide assay was performed by Griess reaction. Some pro-inflammatory cytokines and mediators including TNF-a, IL-1β, IL-6, iNOS, IκB-o. and NF-κB p65 were studied by ELISA, real-time PCR, western blot analysis. Formation of reactive oxygen species (ROS) and expression of CD 14 and TLR4 were evaluated by flow cytometry. To further investigate the anti-inflammatory activity in vivo, we used endotoxemia, sepsis and PMA-induced ear edema animal models.Results:SMLE significantly inhibited the production of NO, TNF-a, IL-1βand IL-6 in LPS-stimulated RAW 264.7 cells. Meanwhile, SMLE also inhibited the LPS-induced cytoplasm IκB-αdegradation and p65 nuclear translocation in RAW 264.7 cells. In addition, SMLE inhibited the production of intracellular ROS and surface expression of CD 14 induced by LPS. In animal model, intraperitoneal administration of SMLE increased the survival rate of endotoxemia and sepsis mice. The topically administration of SMLE significantly inhibited the PMA-induced ear edema.Conclusions:Taken together, SMLE inhibits LPS-induced gene and protein expression of iNOS, TNF-a, IL-1βand IL-6 in macrophages by blocking NF-κB activation and these effects are mediated, at least in part, by inhibiting intracellular ROS generation and surface CD 14 expression. Furthermore, the in vivo anti-inflammatory effect of SMLE suggests a possible therapeutic application of this agent in inflammatory diseases. And these results provide scientific evidence for the traditional Chinese medical practice of S. miltiorrhiza.