绣球藤提取物的抗炎活性及机制

Anti-inflammatory activity and mechanism of Clematis ranunculoides extract

目的探讨绣球藤提取物的抗炎活性及潜在机制。方法使用二甲苯致耳廓肿胀建立急性炎症小鼠模型,以阿司匹林(0.25 g/kg)为阳性对照,1.25、2.5、5 g/kg绣球藤提取物对小鼠耳肿胀度的影响。以植入棉球的方法建立慢性炎症大鼠模型,以阿司匹林(0.17 g/kg)为阳性对照,0.88、1.75、3.5 g/kg绣球藤提取物对大鼠肉芽净重的影响。以脂多糖诱导RAW264.7细胞建立炎症损伤细胞模型,考察12.5、25、50μg/mL绣球藤提取物对细胞上清液中一氧化氮(NO)、前列腺素E2(PGE2)、肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、单核细胞趋化蛋白1(MCP-1)含量,细胞中诱导型一氧化氮合酶(iNOS)、环氧合酶2(COX-2)、p65、磷酸化p65(p-p65)蛋白表达水平,以及p65蛋白核转移的影响。结果5 g/kg的绣球藤提取物可显著降低小鼠的耳肿胀度,1.75、3.5 g/kg的绣球藤提取物可显著降低大鼠的肉芽净重(P<0.05)。12.5、25、50μg/mL绣球藤提取物可显著降低细胞上清液中NO(12.5μg/mL绣球藤提取物除外)、PGE2、TNF-α、IL-6、MCP-1含量,以及细胞中iNOS、COX-2蛋白的相对表达量和p-p65、p65蛋白的相对表达量比值(P<0.05或P<0.01);25、50μg/mL绣球藤提取物可阻断p65蛋白的核转移。结论绣球藤提取物具有良好的抗炎活性,其作用机制可能与抑制核因子κB信号通路激活、下调COX-2和i NOS蛋白的表达、减少炎症因子的释放有关。

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 12.5,25,50μg/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 12.5,25,50μg/mL significantly reduced the contents of NO(except for 12.5μg/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 25,50μg/mL inhibited the translocation of p65 protein to the cell nucleus.CONCLUSIONS C.ranunculoides extract exhibits significant antiinflammatory 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.