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RAW264．7建立炎症细胞模型，加入10μg/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-inflammatory 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-113 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.