丹翘液对脂多糖诱导RAW264.7细胞炎症相关因子的抑制效应分析

doi:10.11843/j.issn.0366-6964.2015.12.023

畜牧兽医学报 ›› 2015, Vol. 46 ›› Issue (12): 2299-2306.doi: 10.11843/j.issn.0366-6964.2015.12.023

丹翘液对脂多糖诱导RAW264.7细胞炎症相关因子的抑制效应分析

魏立琴^1，2，王东升¹，董书伟¹，邝晓娇¹，张世栋^1*，严作廷^1*

(1.中国农业科学院兰州畜牧与兽药研究所/农业部兽用药物创制重点实验室/甘肃省中兽药工程技术研究中心，兰州 730050；2.甘肃农业大学动物医学院，兰州 730070）

收稿日期:2015-04-13 出版日期:2015-12-23 发布日期:2015-12-18
通讯作者: 张世栋（1983-），主要从事药理毒理学研究，E-mail：zhangshidong@caas.cn，Tel：0931-2115262；严作廷（1962-），主要从事中兽医临床和奶牛疾病防治的研究，E-mail：yanzuoting@caas.cn，Tel：0931-2115261
作者简介:魏立琴（1986-），女，甘肃兰州人，硕士，主要从事中兽医临床和奶牛疾病防治的研究，E-mail：24weiliqin@163.com，Tel：0931-2115262
基金资助:
“十二五”国家科技支撑计划项目（2012BAD12B03）；中国农业科学院科技创新工程-奶牛疾病研究

Inhibiting Effect Analysis of Danqiao Liquid on Inflammation-related Factors Induced by LPS in RAW264.7 Cells

WEI Li-qin^1，2，WANG Dong-sheng¹，DONG Shu-wei¹，KUANG Xiao-jiao¹，ZHANG Shi-dong^1* ，YAN Zuo-ting^1*

(1.Lanzhou Institute of Husbandry and Pharmaceutical Science of CAAS/Key Laboratory of Veterinary Pharmaceutics Discovery，Ministry of Agriculture/Engineering & Technology Research Center of Traditional Chinese Veterinary Medicine of Gansu Province，Lanzhou 730050，China；2.College of Veterinary Medicine，Gansu Agricultural University，Lanzhou 730070，China)

Received:2015-04-13 Online:2015-12-23 Published:2015-12-18

摘要/Abstract

摘要：

为研究丹翘液的体外抗炎作用及其抗炎机制，利用LPS诱导巨噬细胞建立炎症模型，MTT法检测不同浓度丹翘液对RAW264.7细胞活力影响；NO测试盒检测丹翘液对LPS诱导的NO释放量的影响；ELISA方法检测丹翘液对LPS诱导的TNF-α、IL-6和PGE₂分泌的影响；RT-PCR方法检测丹翘液对LPS诱导的TNF-α、IL-6、COX-2和iNOS基因转录的影响；Western blot检测对细胞核内NF-κB p65蛋白表达的影响。结果显示丹翘液小于700 μg•mL^-1对细胞无毒性作用；丹翘液各剂量组（100、300、600 μg•mL^-1）能不同程度地抑制LPS诱导的NO、PGE₂、TNF-α和IL-6的分泌；能显著抑制iNOS、COX-2、TNF-α和IL-6基因转录和细胞核内 NF-κB p65蛋白表达。丹翘液的抗炎作用机制可能与抑制NF-κB 通路激活，进而抑制炎症介质和炎性细胞因子的转录和表达有关。

关键词: 丹翘液, 抗炎机制, RAW264.7, LPS, 炎症模型

Abstract:

The aim of the present study was to study the anti-inflammatory effects and possible underlying mechanisms of Danqiao liquid in LPS-stimulated RAW264.7 cells.The cytotoxicity of Danqiao liquid was detected by MTT method.The NO kit assay was adopted to detect the effect of Danqiao liquid on NO release from LPS-induced RAW264.7 cells.ELISA was used to evaluate the production of TNF-α，IL-6 and PGE₂ in LPS-induced RAW264.7 cells.Real-time PCR was used to detect the transcription of COX-2，iNOS，TNF-α and IL-6 in LPS-induced RAW264.7 cells.The protein expression of nuclear NF-κB p65 was detected by Western blot.The result showed that the safe medication range of Danqiao liquid was less than 700 μg•mL^-1.Compared with the LPS model group，Danqiao liquid(100，300，600 μg•mL^-1) could reduce the secretion of NO，PGE₂，TNF-α，and IL-6 in cells induced by LPS，and significantly inhibit the mRNA transcription of iNOS，COX-2，TNF-α，IL-6 and the protein expression of NF-κB p65.In conclusion，this study preliminarily proves the protective effect of Danqiao liquid on LPS-induced RAW264.7 macrophages.Its action mechanism may be related to inhibit NF-κB signal pathway and the genes expressions and secretion of inflammatory mediators and cytokines.

Key words: Danqiao liquid, anti-inflammatory mechanism, RAW264.7, LPS, inflammatory models

中图分类号:

S853.9

魏立琴，王东升，董书伟，邝晓娇，张世栋，严作廷. 丹翘液对脂多糖诱导RAW264.7细胞炎症相关因子的抑制效应分析[J]. 畜牧兽医学报, 2015, 46(12): 2299-2306.

WEI Li-qin，WANG Dong-sheng，DONG Shu-wei，KUANG Xiao-jiao，ZHANG Shi-dong，YAN Zuo-ting. Inhibiting Effect Analysis of Danqiao Liquid on Inflammation-related Factors Induced by LPS in RAW264.7 Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2015, 46(12): 2299-2306.

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丹翘液对脂多糖诱导RAW264.7细胞炎症相关因子的抑制效应分析

Inhibiting Effect Analysis of Danqiao Liquid on Inflammation-related Factors Induced by LPS in RAW264.7 Cells

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