草木犀正丁醇提取物对小鼠巨噬细胞促炎介质的影响

Effect of n-butanol extract from Melilotus on pro-inflammatory factors in murine macrophages

目的研究草木犀正丁醇提取物在体外的抗炎作用。方法通过LPS干预RAW264.7细胞系建立炎症细胞模型,使用1、5和10mg/L3种浓度的草木犀正丁醇提取物对其进行干预。ELISA法检测药物干预前后上清液中TNF-α、IL-1β和IL-6的分泌量,Griess反应测定NO水平;实时荧光定量RT-PCR检测TNF-α、iNOS和COX-2的mRNA表达;Western blot法检测COX-2的蛋白表达水平。结果草木犀正丁醇提取物干预后,上清中TNF-α、IL-1β、IL-6和NO含量与模型组相比均显著降低(P<0.01),并存在剂量依赖关系;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;草木犀正丁醇提取物及DM干预后COX-2蛋白水平明显降低(P<0.01)。结论草木犀正丁醇提取物可以下调TNF-α、IL-1β、IL-6、NO和COX-2等炎症介质的表达,该药物体外抗炎作用可能通过抑制这些炎症介质而产生。

[ Objective ] To investigate the anti-inflammatory mechanism of n-butanol extract from Melilotus suaveolens Ledeb（MSL） in vitro. [ Method] The cell model was established by lipopolysaecharide（LPS） induction on the RAW264.7 maerophages cell line. ELISA assay was used to measure the secretion of TNF-α, IL-1β and IL-6 in the supematant of MSL treated and untreated macrophages. 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 the controls （P 〈0.01）. We also found signifieant 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 n-butanol extract from Melilotus suaveolens Ledeb could inhibit inflammation by down-regulating the expression and secretion of pro-inflammatory factors.