Tao Jun-Yan, Zheng Guo-Hua, Zhao Lei, Wu Jian-Guo, Zhang Xiao-Yu, Zhang Shu-Ling, Huang Zhi-Jun, Xiong Fu-Liang, Li Chong-Ming
State Key Laboratory of Virology, College of Life Sciences, Chinese-French Liver Disease Research Institute of Wuhan University at Zhongnan Hospital, Wuhan University, Wuhan, 430072, PR China.
J Ethnopharmacol. 2009 May 4;123(1):97-105. doi: 10.1016/j.jep.2009.02.024. Epub 2009 Mar 3.
This paper aimed to elucidate the anti-inflammatory effects of EtOAc fraction prepared from Melilotus suaveolens Ledeb ethanol extract with a cellular model of LPS-stimulated RAW 264.7 cell.
Some key pro-inflammatory cytokines and mediators including IL-1 beta, IL-6, NO, iNOS, COX-2 and TNF-alpha, two important anti-inflammatory cytokines and mediators IL-10 and HO-1, I-kappaB and NF-kappaB were studied by sandwich ELISA, real-time PCR, western blot analysis and immunocytochemistry. At last a HPLC fingerprint was taken to evaluate the fraction.
The EtOAc fraction could significantly inhibit the production of IL-1 beta, IL-6, NO, TNF-alpha, COX-2 in LPS-stimulated cell than that of single LPS-stimulated cell (p<0.01 or p<0.05), and the extract could increase the production of IL-10 and HO-1 than that of single LPS intervention cell (p<0.01 or p<0.05). Meanwhile, the extract also could inhibit the production of NF-kappaB compared to single LPS-stimulated cell. All the results showed that the extract had a good anti-inflammatory effect on LPS-stimulated RAW264.7 cell.
Taken together, the anti-inflammatory actions of M. suaveolens Ledeb EtOAc fraction might be due to the down-regulation of IL-1 beta, IL-6, NO, TNF-alpha and COX-2 via the suppression of NF-kappaB activation, and another pathway was up regulating the production of IL-10 and HO-1. Meanwhile, the EtOAc fraction might be further studied to isolate the active anti-inflammatory ingredients besides coumarin.
本文旨在通过脂多糖（LPS）刺激的RAW 264.7细胞的细胞模型，阐明草木犀乙醇提取物的乙酸乙酯部位的抗炎作用。
采用夹心酶联免疫吸附测定（ELISA）、实时荧光定量聚合酶链反应（PCR）、蛋白质免疫印迹分析和免疫细胞化学等方法，研究了一些关键的促炎细胞因子和介质，包括白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、一氧化氮（NO）、诱导型一氧化氮合酶（iNOS）、环氧化酶-2（COX-2）和肿瘤坏死因子-α（TNF-α），以及两种重要的抗炎细胞因子和介质白细胞介素-10（IL-10）和血红素氧合酶-1（HO-1）、核因子κB抑制蛋白（I-κB）和核因子κB（NF-κB）。最后采用高效液相色谱（HPLC）指纹图谱对该部位进行评价。
与单纯LPS刺激的细胞相比，乙酸乙酯部位能显著抑制LPS刺激细胞中IL-1β、IL-6、NO、TNF-α、COX-2的产生（p<0.01或p<0.05），且该提取物能增加IL-10和HO-1的产生，与单纯LPS干预的细胞相比（p<0.01或p<0.05）。同时，与单纯LPS刺激的细胞相比，该提取物还能抑制NF-κB的产生。所有结果表明该提取物对LPS刺激的RAW264.7细胞具有良好的抗炎作用。
综上所述，草木犀乙酸乙酯部位的抗炎作用可能是通过抑制NF-κB活化，下调IL-1β、IL-6、NO、TNF-α和COX-2的表达，另一条途径是上调IL-10和HO-1的产生。同时，除香豆素外，乙酸乙酯部位可能还需进一步研究以分离出活性抗炎成分。
