摘要
目的探讨白桦脂醇对Aβ25-35诱导的小胶质细胞炎症反应的保护作用。方法MTT法预实验证实10μmol/L Aβ25-35的浓度对细胞活力无影响,但是影响炎症因子分泌,遂以10μmol/L Aβ25-35作用于BV2细胞,实验随机分为对照组、模型组、白桦脂醇5、10和20μmol/L组,采用MTT法检测细胞的存活率、Western blotting检测NF-κB信号通路蛋白(NF-κB P65、IκBα、p-NF-κB P65、p-IκBα)表达水平及炎症诱导酶环氧合酶2(COX-2)、诱导型一氧化氮合成酶(iNOS)的水平,以及炎症因子白细胞介素1β(IL-1β),肿瘤坏死因子α(TNF-α)及IL-10水平。结果Aβ25-35浓度为10μmol/L时,对BV2细胞存活率无影响。白桦脂醇浓度在0~20μmol/L时低毒性、细胞活力无变化。白桦脂醇预处理组IκBα、NF-κB P65的蛋白表达水平升高,p-NF-κB P65和p-IκBα的蛋白表达水平显著降低,COX-2、iNOS的水平显著降低,促炎因子IL-1β、TNF-α的表达降低,而抑炎因子IL-10的表达升高。结论白桦脂醇对Aβ25-35诱导的BV2细胞的炎症反应有抑制作用,通过NF-κB信号通路,抑制iNOS及COX-2蛋白表达,并抑制TNF-α和IL-1β水平,提高IL-10水平。
Microglia mediated inflammatory response plays an important role in the course of Alzheimer’s disease,while betulin can inhibit the inflammatory response to a certain extent.Therefore,this study was designed to investigate the effect and mechanism of betulin on the inflammatory response of BV2 cells induced by Aβ25-35 via NF-κB signaling pathway.BV2 cells were randomly divided into control group,model group,betulin 5,10,and 20μmol/L groups.MTT assay was used to measure cell viability;Western blot was employed to detect the expression of NF-κB signal pathway proteins(NF-κB P65,IκBα,p-NF-κB P65,p-IκBα)and inflammatory inducing enzyme cyclooxygenase.Western blot was also used to detect the activation level of enzyme 2(COX-2)and inducible nitric oxide synthase(iNOS),the levels of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and IL-10.Data showed that low concentration of Aβ25-35(10μmol/L)did not affect the survival rate of BV2 cells.Betulin at 0~20μmol/L was low toxicity and did not affect cell viability.In betulin groups,the protein expression levels of IκBαand NF-κB P65 increased,p-NF-κB P65 and p-IκBαdecreased significantly,COX-2 and iNOS activation levels decreased significantly,the expression of pro-inflammatory factors IL-1βand TNF-αdecreased,while the expression of anti-inflammatory factors IL-10 increased as compared with control group.In conclusion,betulin can inhibit the inflammatory response of BV2 cells induced by Aβ25-35,inhibit the expression of iNOS and COX-2 protein,inhibit the level of TNF-αand IL-1β,and improve the level of IL-10 through NF-κB signal pathway.
作者
傅增辉
姜岩
刘晶
林再红
杜姝
张广萍
金艳
陈团团
FU Zenghui;JIANG Yan;LIU Jing;LIN Zaihong;DU Shu;ZHANG Guangping;JIN Yan;CHEN Tuantan(Department of Neurology,Third Affiliated Hospital of Qiqihar Medical University,Qiqihar 161002,China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2020年第9期784-789,共6页
Immunological Journal
基金
黑龙江省教育厅基本科研业务费基础研究项目(2017-KYYWF-0700)。
