木犀草素降低大肠埃希菌对呼吸道上皮细胞感染的作用机制研究  

Effect of luteolin on reducing the infection of Escherichia coli in respiratory epithelial cells

作　　者：黄志勇 孙凤军[2] 袁慊 王瑜[2] 熊丽蓉[2] 冯伟[2] HUANG Zhi-yong;SUN Feng-jun;YUAN Qian;WANG Yu;XIONG Li-rong;FENG Wei(Department of Spine and Joint,The First Naval Hospital of Southern Theater Command,Guangdong Zhanjiang524005,China;Department of Pharmacy,Southwest Hospital,Army Medical University(Third Military Medical University),Chongqing 400038,China)

机构地区：[1]南部战区海军第一医院脊柱关节科,广东湛江524005 [2]陆军军医大学(第三军医大学)西南医院药剂科,重庆400038

出　　处：《中国医院药学杂志》2020年第7期781-785,共5页Chinese Journal of Hospital Pharmacy

基　　金：重庆市应用开发计划项目(编号:cstc2014yykfA110021)。

摘　　要：目的:探讨亚抑菌浓度(sub-minimal inhibitory concentration,亚-MIC)木犀草素降低临床分离大肠埃希菌对呼吸道上皮细胞HBE感染的作用机制,为临床抗感染的治疗提供新的思路。方法:采用微量肉汤稀释法检测木犀草素对大肠埃希菌临床株E367的MIC值,采用多功能酶标仪测定亚-MIC木犀草素对细菌生长的影响,采用CCK-8法测定亚-MIC木犀草素处理大肠埃希菌对HBE细胞毒性的影响,采用ELISA试剂盒测定细胞炎症因子白细胞介素(interleukin,IL)-6、IL-8及肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,采用Western Blot测定p-p65/p65、p-IκBα/IκBα蛋白表达水平。结果:1/4 MIC-1/32 MIC木犀草素对大肠埃希菌的生长没有影响,且能剂量依赖性地降低大肠埃希菌对呼吸道上皮细胞的毒性。大肠埃希菌感染细胞后其炎症因子IL-6、IL-8、TNF-α水平和p-p65/p65、p-IκBα/IκBα蛋白表达水平均显著增高。与细菌处理组相比,木犀草素和大肠埃希菌共同处理细胞后其IL-6、TNF-α水平及p-p65/p65、p-IκBα/IκBα蛋白表达水平显著降低,而细胞因子IL-8水平几乎不变。结论:木犀草素可能通过免疫调节抑制大肠埃希菌对呼吸道上皮细胞的感染作用。OBJECTIVE To investigate the mechanism of sub-minimal inhibitory concentration(sub-MIC)luteolin on reducing the infection of Escherichia coli(E.coli)in respiratory epithelial cells HBE,and provide new ideas for the treatment of clinical anti-infection.METHODS The MIC of luteolin on E.coli isolate E367 was detected by microdilution broth method.The effect of sub-MIC luteolin on bacterial growth was determined by automatic microplate reader.The effect of luteolin on the cytotoxicity of E.coli in respiratory epithelial cells was determined by CCK-8 method.The levels of interleukin(IL)-6,IL-8 and tumor necrosis factor-α(TNF-α)were determined by ELISA kit.The expression levels of p-p65/p65,p-IκBα/IκBαproteins were determined by Western Blot.RESULTS 1/4 MIC-1/32 MIC luteolin has no effect on the growth of E.coli isolate E367 and could reduce the cytotoxicity of E.coli in respiratory epithelial cells in a dose-dependent manner.The production of cytokines IL-6,IL-8,TNF-αand the levels of p-p65/p65,p-IκBα/IκBαprotein expression were significantly increased in cells infected with E.coli.Compared with the bacterial treatment group,the levels of IL-6,TNF-αproduction and p-p65/p65、p-IκBα/IκBαprotein expression were significantly decreased when the cells were treated with both luteolin and E.coli,while the level of cytokine IL-8 was almost unchanged.CONCLUSION Luteolin may inhibit the infection of E.coli in respiratory epithelial cells through immune regulation.

关 键 词：木犀草素 大肠埃希菌 呼吸道上皮细胞 亚抑菌浓度 细胞毒性 

分 类 号：R969[医药卫生—药理学]