异槲皮苷对脂多糖诱导RAW264.7细胞产生炎症因子的调控  

Regulation Effects of Isoquercitrin on Inflammatory in LPS-induced RAW264.7 Cell

作　　者：刘天旭[1] 李娟[1] 蒋国君[1] 董晓敏[2] 朱钊铭 黄桂红[1,3] 

机构地区：[1]桂林医学院临桂临床医学院,桂林541000 [2]桂林医学院附属医院药学部,桂林541000 [3]桂林医学院第二附属医院药学部,桂林541000

出　　处：《医药导报》2017年第6期601-605,共5页Herald of Medicine

基　　金：国家自然科学基金资助项目(81260661);广西自然科学基金项目资助(2012GXNSFAA053079);桂林市科学研究与技术开发项目(20140120-1-9);广西肝损伤与修复分子医学重点实验室科研基金资助项目(15-140-46-14)

摘　　要：目的探讨异槲皮苷对脂多糖(LPS)诱导RAW264.7细胞产生的炎症因子的调控作用。方法噻唑蓝(MTT)法检测异槲皮苷对细胞生长的抑制率;酶联免疫吸附测定(ELISA)法检测培养基中肿瘤坏死因子α(TNF-α)含量,一氧化氮(NO)试剂盒检测NO含量;Western blotting检测i NOS和环氧化酶-2(COX-2)蛋白的表达。结果异槲皮苷对细胞的半数抑制浓度为65.73μmol·L^(-1);LPS对细胞无抑制作用;与LPS组比较,异槲皮苷(20,10μmol·L^(-1))可使TNF-α分泌降低至74.80%,60.57%,且对TNF-α分泌的抑制作用,与剂量减少呈正相关;一氧化氮试剂盒检测结果显示异槲皮苷(20,10μmol·L^(-1))可以抑制NO的分泌,NO的分泌量降低至79.34%,68.81%(P<0.05);Western blotting实验结果表明:异槲皮苷(20,15,10μmol·L^(-1))可降低i NOS和COX-2蛋白的表达(P<0.05)。结论异槲皮苷具有抗炎作用,其抗炎分子机制可能与抑制TNF-α、NO、iNOS和COX-2的产生有关,当异槲皮苷浓度为10μmol·L^(-1)时抗炎作用最强。Objective To investigate the influence of isoquercitrin on the inflammatory factors in LPS-induced RAW264. 7 cells. Methods MTT method was used to detect inhibition ratio of RAW 264. 7 cells induced by isoquercitrin. The level of TNF-α in culture medium was measured by ELISA. Nitric oxide（ NO） was detected by Nitrate Assay Kit. Western blotting was used to investigate the influence on the productions of inducible nitric oxide synthase（ i NOS） and cyclooxygenase-2（ COX-2）. Results The half inhibitory concentration（ IC50） of isoquercitrin was 65. 73 μmol·L^（-1）. LPS had no inhibitory effect on the cells. Compared with LPS group,the level of TNF-α was decreased to 74. 80% and 60. 57% in isoquercitrin（ 20,10 μmol·L^（-1）） groups in a dose-dependent manner. The results measured by Nitrate Assay Kit revealed that isoquercitrin（ 20,10 μmol·L^（-1）） could suppress production of NO,the level of NO decreased to 79. 34% and 68. 81%（ P〈 0. 05）. The Western blotting results showed that isoquercitrin（ 20,15,10 μmol·L^（-1）） inhibited the productions of i NOS and COX-2（ P〈 0. 05）.Conclusion Isoquercitrin has anti-inflammatory effects by inhibiting the productions of TNF-α,NO,i NOS and COX-2,and the most effective dose for the inhibition is 10 μmol·L^（-1）.

关 键 词：异槲皮苷 RAW264.7细胞 炎症因子 肿瘤坏死因子α 

分 类 号：R965[医药卫生—药理学]