草木犀石油醚提取物对小鼠巨噬细胞促炎介质的影响

目的 研究草木犀石油醚提取物在体外的抗炎作用.方法 采用小鼠巨噬细胞系RAW 264.7建立炎症细胞模型,加入10 μg/L的LPS培养液和不同浓度的草木犀石油醚提取物进行干预.ELISA法检测上清液中TNF-α, IL-1β, IL-6和NO的分泌量;实时荧光定量RT-PCR检测TNF-α, iNOS 和 COX-2的 mRNA表达;Western 印迹法检测COX-2 蛋白的表达.结果 草木犀提取物干预后细胞所分泌的炎性介质(TNF-α, IL-1β, IL-6和NO)与模型组相比均显著降低(P＜0.01),并存在剂量依赖关系;RT-PCR结果显示干预后细胞TNF-α,iNOS和COX-2的mRNA表达水平显著降低(P＜0.01),也存在剂量依赖关系;Western印迹结果显示草木犀石油醚提取物及地塞米松干预后COX-2蛋白水平明显降低(P＜0.01).结论 草木犀的石油醚提取物通过下调LPS诱导的巨噬细胞表达炎性介质而发挥其体外抗炎作用, 且其下调作用呈剂量依赖性.

Effect of Petroleum Ether Extract from Melilotus on Pro-inflam matory Mediators in Murine Macrophages

Objective To study anti-inflammatory mechanism of petroleum ether extract from Melilotus suaveolens Ledeb in vitro.Method Inflammatory cell model was established by LPS acting on the RAW264.7 cell line. The secretion of TNF-α, IL-1β and IL-6 in supernatant was detected by ELISA. The mRNA expression of TNF-α, iNOS, COX-2 and the protein expression of COX-2 were detected by real-time PCR and Western blot respectively.Results The secretion levels of TNF-α, IL-1β, IL-6 and NO were significantly lower in the supernatant of MSL treated macrophages than those in controls (P<0.01). We also found significant differences among the mRNA expression of TNF-α, iNOS and COX-2 between the supernatant of MSL treated and untreated macrophages, as well as the protein expression of COX-2 (P<0.01).Conclusion The petroleum ether extract from Melilotus suaveolens Ledeb can significantly resist inflammation by inhibiting expression and secretion of pro-inflammatory mediators in murine macrophages treated by LPS.