Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The anti...Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The antiinflammatory effects of EFPF were evaluated using the xylene-induced mouse ear edema and carrageenan-induced rat paw edema models in vivo. In vitro, RAW264.7 cells were exposed to 0–800 μg/m L EFPF and the cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay. Then cells were treated with different concentrations of EFPF（100–400 μg/m L） and stimulated with lipopolysaccharide（LPS, 1 μg/m L） for 24 h. The supernatant was analyzed for nitric oxide（NO） using the Griess reagent, and the levels of inflammatory mediators and cytokines were determined using enzyme-linked immunosorbent assays for prostaglandin E2（PGE2）, tumor necrosis factor α（TNF-α）, interleukin（IL） 6, and IL-10. The protein expressions of inducible NO synthase（i NOS）, cyclooxygenase-2（COX-2）, nuclear factor κB（NF-κB）, and mitogen-activated protein kinases（MAPKs） including extracellular signal-regulated kinase（ERK）, c-Jun N-terminal kinase（JNK）, and p38 MAPK were examined by Western blot. Results：Compared with the control group, EFPF significantly reduced mouse ear edema and rat paw edema rate（P〈0.05 or P〈0.01）. Compared with the LPS group, EFPF significantly inhibited the LPS-stimulated production of NO, PGE2, TNF-α and IL-6（P〈0.05 or P〈0.01）, and increased the IL-10 production（P〈0.05）. EFPF also significantly inhibited LPS-induced protein expressions of i NOS and COX-2, suppressed the phosphorylation and degradation of inhibitor of NF-κB-α, decreased p65 level, and inhibited the phosphorylation of p38, ERK1/2 and JNK（P〈0.05 or P〈0.01）. Conclusion：EFPF exerted anti-inflammatory effect by reducing protein expressions of i NOS and COX-2 and the production of the inflammation factors, including TNF-α, IL-6, NO and PGE2, mainly through inhibition of LPS-mediated stimulation of NF-κB and MAPK signaling pathways.展开更多
基金Supported by the National Natural Science Foundation of China(No.81460641,No.81660691)
文摘Objective：To determine the anti-inflammatory effects of an ethanol fraction of Periploca forrestii Schltr.（EFPF） and to investigate the potential mechanisms underlying in vivo and in vitro models. Methods：The antiinflammatory effects of EFPF were evaluated using the xylene-induced mouse ear edema and carrageenan-induced rat paw edema models in vivo. In vitro, RAW264.7 cells were exposed to 0–800 μg/m L EFPF and the cell viability was determined by 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide（MTT） assay. Then cells were treated with different concentrations of EFPF（100–400 μg/m L） and stimulated with lipopolysaccharide（LPS, 1 μg/m L） for 24 h. The supernatant was analyzed for nitric oxide（NO） using the Griess reagent, and the levels of inflammatory mediators and cytokines were determined using enzyme-linked immunosorbent assays for prostaglandin E2（PGE2）, tumor necrosis factor α（TNF-α）, interleukin（IL） 6, and IL-10. The protein expressions of inducible NO synthase（i NOS）, cyclooxygenase-2（COX-2）, nuclear factor κB（NF-κB）, and mitogen-activated protein kinases（MAPKs） including extracellular signal-regulated kinase（ERK）, c-Jun N-terminal kinase（JNK）, and p38 MAPK were examined by Western blot. Results：Compared with the control group, EFPF significantly reduced mouse ear edema and rat paw edema rate（P〈0.05 or P〈0.01）. Compared with the LPS group, EFPF significantly inhibited the LPS-stimulated production of NO, PGE2, TNF-α and IL-6（P〈0.05 or P〈0.01）, and increased the IL-10 production（P〈0.05）. EFPF also significantly inhibited LPS-induced protein expressions of i NOS and COX-2, suppressed the phosphorylation and degradation of inhibitor of NF-κB-α, decreased p65 level, and inhibited the phosphorylation of p38, ERK1/2 and JNK（P〈0.05 or P〈0.01）. Conclusion：EFPF exerted anti-inflammatory effect by reducing protein expressions of i NOS and COX-2 and the production of the inflammation factors, including TNF-α, IL-6, NO and PGE2, mainly through inhibition of LPS-mediated stimulation of NF-κB and MAPK signaling pathways.
