Anti-inflammatory activity and mechanism of Clematis ranunculoides extract
OBJECTIVE To investigate the anti-inflammatory activity and potential mechanism of Clematis ranunculoides extract.METHODS The ear swelling was induced by xylene to establish an acute inflammation model of mice;using aspirin(0.25 g/kg)as a positive control,the effects of 1.25,2.5,5 g/kg C.ranunculoides extract on the degree of ear swelling were investigated.The chronic inflammation model of rats was also established by implanting cotton balls;using aspirin(0.17 g/kg)as a positive control,the effects of 0.88,1.75,3.5 g/kg C.ranunculoides extract on the net weight of granulomas were investigated.Furthermore,RAW264.7 cells were induced by lipopolysaccharide to establish an inflammatory injury model;the effects of 12.5,25,50 μg/mL C.ranunculoides extract on the contents of nitric oxide(NO),prostaglandin E2(PGE2),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6)and monocyte chemotactic protein-1(MCP-1)in the cell supernatant,the protein expressions of inducible nitric oxide synthase(iNOS),cyclooxygenase-2(COX-2),p65 and phosphorylated p65(p-p65)in cells as well as nuclear translocation of p65 protein were assessed.RESULTS C.ranunculoides extract with 5 g/kg significantly relieved ear swelling in mice,and C.ranunculoides extract with 1.75,3.5 g/kg significantly decreased the net weight of granulomas in rats(P＜0.05).C.ranunculoides extract with 12.5,25,50 μg/mL significantly reduced the contents of NO(except for 12.5 μg/mL C.ranunculoides extract),PGE2,TNF-α,IL-6 and MCP-1 in the cell supernatant,as well as the relative expressions of iNOS and COX-2 protein,and relative expression ratio of p-p65 and p65 protein(P＜0.05 or P＜0.01);C.ranunculoides extract with 25,50 μg/mL inhibited the translocation of p65 protein to the cell nucleus.CONCLUSIONS C.ranunculoides extract exhibits significant anti-inflammatory activity,the mechanism of which may be attributed to the inhibition of the activation of nuclear factor-κB signaling pathway,down-regulation of COX-2 and iNOS protein expression,and the reduction of inflammatory cytokines release.
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