RESUMO
Objective:To investigate the effect of epigallocatechin-3-gallate ( EGCG) on the expression of pro-inflammatory cytokines in murine bone marrow-derived macrophages ( BMDM) induced by IFN-γ/LPS.Methods:The bone marrow cells were isolated from 6-8 weeks C57BL/6 mice,which cultured in RPMI1640 medium with 10%FBS and stimulated with 100 ng/ml M-CSF in vitro,and then exposed to 50 ng/ml IFN-γand 1 μg/ml LPS with various concentrations of EGCG (12.5-50 μmol/L) .The expression of pro-inflammatory factors,IL-1β,TNF-αand iNOS were detected,in murine myeloid macrophages stimulated by IFN-γ/LPS with qRT-PCR and ELISA.Results:IFN-γ/LPS remarkably up-regulated the expression levels of inflammatory cytokines ,IL-1β,TNF-αand iNOS,but EGCG effectively repressed these cytokines expression in IFN-γ/LPS-stimulated murine myeloid macrophages , in dose-dependent manner.Conclusion:EGCG attenuates the pro-inflammatory phenotype of murine myeloid macrophages stimulated with IFN-γ/LPS.
RESUMO
Hydrophobic interaction chromatography was used to separate correctly refolded and mis-refolded consensus interferon. The effects of ligand types, salt concentration, pH and flow rate were investigated. The best result could be obtained by using Butyl Sepharose 4 Fast Flow, 0.8 mol/L of ammonium sulfate, pH 8.3 and 90cm/h of linear flow rate. Reverse-phase HPLC analysis showed the purity of the pooled fraction was as high as 99.6%. The specific activity of purified consensus interferon was 2.3 x 10(9) IU/mg, The mass recovery of targeth protein was 36.7%.