[栀子苷对脂多糖诱导RAW264.7巨噬细胞系TLR4-NF-κB通路激活的影响]
[Effect of geniposide on LPS-induced activation of TLR4-NF-κB pathway in RAW264.7 macrophage cell line].
作者信息
Huang Lihua, Wang Chunjie, Naren Gaowa, Aori Gele
机构信息
College of Veterinarian, Inner Mongolia Agricultural University, Huhhot (Huhehaote) 010010, China.
出版信息
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2013 Oct;29(10):1012-4.
OBJECTIVE
To study the anti-inflammatory mechanism of geniposide and observe the effect of geniposide on the expression of Toll-like receptor 4 (TLR4), the activity of NF-κB, and the release of pro-inflammatory cytokines- TNF-α, IL-1, and IL-6-in the RAW264.7 macrophages treated with lipopolysaccharide (LPS).
METHODS
There were three experimental groups, including the control group, LPS group and LPS combined with geniposide group in this study. RAW264.7 macrophage cells were treated with LPS to induce cellular inflammation. Cell proliferation was measured by CCK-8. The concentrations of TNF-α, IL-1, and IL-6 in cell culture media were measured by ELISA. mRNA levels of TLR4 and P65 were examined by real-time PCR. The protein levels of p-IκB, P65, p-P65 and TLR4 were detected by Western blotting.
RESULTS
Geniposide had no effect on cell proliferation. However, geniposide down-regulated the expression of TNF-α, IL-1, and IL-6, and also inhibited the expression of TLR4 and the activity of NF-κB.
CONCLUSION
Geniposide exerts its anti-inflammatory effect through inhibiting the activity of NF-κB in the TLR4-NF-κB pathway in macrophages.
目的
研究栀子苷的抗炎机制，观察栀子苷对脂多糖（LPS）处理的RAW264.7巨噬细胞中Toll样受体4（TLR4）表达、核因子κB（NF-κB）活性及促炎细胞因子肿瘤坏死因子-α（TNF-α）、白细胞介素-1（IL-1）和白细胞介素-6（IL-6）释放的影响。
方法
本研究设有三个实验组，即对照组、LPS组和LPS联合栀子苷组。用LPS处理RAW264.7巨噬细胞以诱导细胞炎症。采用CCK-8法检测细胞增殖。用酶联免疫吸附测定法（ELISA）检测细胞培养基中TNF-α、IL-1和IL-6的浓度。通过实时聚合酶链反应（real-time PCR）检测TLR4和P65的mRNA水平。用蛋白质印迹法检测磷酸化IκB（p-IκB）、P65、磷酸化P65（p-P65）和TLR4的蛋白水平。
结果
栀子苷对细胞增殖无影响。然而，栀子苷下调了TNF-α、IL-1和IL-6的表达，同时抑制了TLR4的表达和NF-κB的活性。
结论
栀子苷通过抑制巨噬细胞中TLR4-NF-κB途径中NF-κB的活性发挥抗炎作用。
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