洋葱槲皮素对巨噬细胞HO-1表达影响及相关信号通路的研究  

Effect of onion quercetin on HO-1 expression in macrophages and related pathway

作　　者：张敏 王筱婧[2] ZHANG Min;WANG Xiaojing(Department of Reproductive Immunology, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai 201204, China;International Cooperation Department, Chinese Pharmaceutical Association, Beijing 100050, China)

机构地区：[1]同济大学附属第一妇婴保健院生殖免疫科,上海201204 [2]中国药学会国际合作部,北京100050

出　　处：《免疫学杂志》2019年第8期732-736,共5页Immunological Journal

摘　　要：目的洋葱槲皮素具有抗炎作用,但具体机制尚不明确。本研究主要探讨其对RAW 264.7巨噬细胞HO-1表达影响及相关信号通路的作用。方法采用Griess法检测NO水平;用Western blot法检测HO-1、Nrf2、Keap1的表达水平;免疫共沉淀法检测Nrf2和Keap1相互作用;用双荧光素酶报告检测考察Nrf2与ARE的结合水平。结果 Western blot结果显示洋葱槲皮素能显著提高LPS诱导的HO-1表达水平,增加Nrf2的表达水平,且降低Keap1的表达水平。免疫共沉淀法结果表明,无论是否存在LPS诱导,洋葱槲皮素均可显著降低Keap1/Nrf2比值。双荧光素酶报告实验的结果表明,洋葱槲皮素能增强ARE介导的荧光素酶活性。SnPP实验结果表明,SnPP能减弱洋葱槲皮素对NO、iNOS和NF-κB的抑制作用。结论洋葱槲皮素可能通过诱导HO-1的表达产生抗炎作用,这种诱导作用可能与Keap1/Nrf2/ARE信号通路有关。Onion quercetin has anti-inflammatory effect, but its mechanism is still unclear. This study was to investigate its effect on HO-1 expression in RAW 264.7 macrophages and its related pathway. The levels of HO-1,Nrf2, and Keap1 were detected by Western blot. The interaction between Nrf2 and Keap1 was detected by coimmunoprecipitation. The binding level of Nrf2 and ARE was detected by double luciferase report. Western blot showed that onion quercetin could significantly increase the expression level of HO-1 and Nrf2 induced by LPS, and decrease the level of Keap1. The results of co-immunoprecipitation showed that onion quercetin could significantly reduce the Keap1/Nrf2 ratio regardless of the LPS induction. The results of double luciferase report test showed that onion quercetin could enhance the ARE-mediated luciferase activities. The results of SnPP test showed that SnPP could attenuate the inhibitory effects of onion quercetin on NO, iNOS and NF-κB. The anti-inflammatory effect of onion quercetin could be attributed to the induction of the HO-1 expression. This induction may be related to Keap1/Nrf2/ARE pathway.

关 键 词：洋葱槲皮素 抗炎 HO-1 Keap1/Nrf2/ARE信号通路 

分 类 号：R392.11[医药卫生—免疫学]