Active constituents and anti-inflammatory mechanism of Fangji Fuling Decoction

AIM To study the active constituents and anti-inflammatory mechanism of Fangji Fuling Decoction(Stephaniae tetrandrae Radix,Astragali Radix,Cinnamomi Ramulus,Poria,Glycyrrhizae Radix et Rhizoma).METHODS Water-soluble and ethanolic extract was prepared and constituents extracted by petroleum ether,ethyl acetate,n-butanol,water were separated.Inflammatory cell model was developed by the combination of IFN-γ with LPS stimulating RAW 264.7 macrophage.Griess reaction was used for nitric oxide accumulation,MTT assay for cell proliferation ability,FRAP assay for total antioxidant capacity,and Western blot for protein expression.RESULTS Compared with other constituents,ethyl acetate fraction from Fangji Fuling Decoction ethanol(EAFD) extract exhibited the strongest suppressing effect on nitric oxide formation in dose-dependent manner and no cytotoxicity in resting macrophage cells.The protein expression of inducible nitric oxide synthase(iNOS) and cyclooxygenase-2(COX-2) were inhibited by EAFD,at the same time increased total antioxidant capacity.CONCLUSION These results suggest that EAFD may provide potential anti-inflammatory effect through inhibiting iNOS and COX-2 protein expression and increase the total antioxidant capacity.