摘要
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、50 μg/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、50 μg/mL绣球藤提取物可显著降低细胞上清液中NO(12.5 μg/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、50 μg/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iNOS蛋白的表达、减少炎症因子的释放有关.
Abstract
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.
基金项目
云南省创新引导与科技型企业培育计划项目(202004AR040008)
维普
万方数据