Amelioration of Danhong injection on the lipopolysaccharide-stimulated systemic acute inflammatory reaction via multi-target strategy.

ETHNOPHARMACOLOGICAL RELEVANCE: Systemic inflammatory response syndrome (SIRS), leading to dire consequences, is a serious and fatal disease in clinic. Danhong injection (DHI), one of the most popular medications for coronary heart disease and cerebral ischemia, plays pharmacological actions through inhibiting local inflammation. Nevertheless, the anti-inflammatory effect of DHI has not been reported before and has not been fully clarified. AIM OF THE STUDY: In this study, a model of systemic acute inflammatory reaction was induced by lipopolysaccharide (LPS) to investigate whether DHI could be applied to SIRS through the anti-inflammatory effect.
MATERIAL AND METHODS: The anti-inflammatory effect of DHI in vivo was evaluated in ICR mice pretreated intraperitoneally (i.p.) with LPS (1mg/kg) and the serum, liver and kidney were collected. Interleukin (IL)-6, tumor necrosis factor (TNF)-α and monocyte chemotactic protein (MCP-1) in serum were measured by enzyme-linked immunosorbent assay (ELISA) and the mRNA expressions of inducible NO synthase (iNOS), IL-6, interleukin (IL)-1β, MCP-1 in mice liver and kidney were analyzed by quantitative real-time reverse-transcription polymerase chain reaction (real-time RT-PCR). Meanwhile, Proteome profiler array was used to screen the acute phase proteins, cytokines and chemokines activated in the acute inflammation. The inflammatory model of macrophages stimulated by LPS (0.2μg/mL) was used to evaluate the anti-inflammatory mechanism of DHI in vitro. The secretion of nitric oxide (NO) was measured by the Griess reagent system. The productions of prostaglandin E2 (PGE2), IL-6, TNF-α and MCP-1 were detected using ELISA, and the protein expression of cyclooxygenase (COX)-2 was determined by cell-based ELISA. As well, the mRNA expressions of these inflammatory factors were detected by real-time RT-PCR.
RESULTS: DHI could attenuate the inflammatory reaction via decreasing 20 cytokines and acute phase proteins analyzed by Proteome profile array in serum. The secretions of IL-6, TNF-α and MCP-1 in serum were coincidence with the result of Proteome profile array. Meanwhile, the mRNA expressions of iNOS, IL-6, IL-1β, MCP-1 in mice liver and kidney were significantly reduced by DHI. Experiments performed in vitro further revealed that the productions of NO, PGE2 and the mRNA expressions of iNOS, COX-2 were notably inhibited by DHI. Cell-based ELISA revealed that the COX-2 protein expression was diminished by DHI. The results of ELISA demonstrated that DHI significantly down-regulated the protein productions of IL-6 and MCP-1. Furthermore, the mRNA expressions of iNOS, COX-2, TNF-α, IL-1β, IL-6 and MCP-1 analyzed by real-time RT-PCR were suppressed by DHI.
CONCLUSIONS: These results demonstrate that DHI exerts the protective effect through inhibiting the expressions of iNOS, COX-2, IL-1β, IL-6, MCP-1 and TNF-α, which elucidate that DHI may be a strongly multi-target Chinese medicine injection on improving the inflammatory diseases.