甜叶悬钩子苷通过NF-κB和MAPK信号通路抑制小鼠神经炎症细胞模型炎症反应  

Rubusoside inhibits inflammatory response in a mouse neuroinflammato⁃ry cell model via NF-κB and MAPK signaling pathways

作　　者：杨爽 董明鑫[2] 姜新 邴艳华[1] 王燕 张剑旭[2] 孙成彪 岳灿灿 许娜 刘文森 YANG Shuang;DONG Mingxin;JIANG Xin;BING Yanhua;WANG Yan;ZHANG Jianxu;SUN Chengbiao;YUE Cancan;XU Na;LIU Wensen(Department of Physiology and Pathophysiology,Yanbian University Medical School,Yanji 133002,China;Changchun Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Changchun 130000,China;Jilin Medical Col-lege,Jilin132000,China)

机构地区：[1]延边大学医学院生理学与病理生理学教研室,吉林延吉133002 [2]中国农业科学院长春兽医研究所,吉林长春130000 [3]吉林医药学院,吉林吉林132000

出　　处：《中国病理生理杂志》2023年第4期616-622,共7页Chinese Journal of Pathophysiology

基　　金：吉林省教育厅科研课题(No.JJKH20210503KJ)。

摘　　要：目的:通过体外实验探究甜叶悬钩子苷(RUB)对脂多糖(LPS)诱导小鼠BV-2小胶质细胞神经炎症反应的影响及改善机制。方法:采用CCK-8法检测RUB对BV-2细胞活力的影响;使用LPS诱导BV-2小胶质细胞建立神经炎症细胞模型,用不同浓度RUB进行干预,将BV-2细胞分为对照组、LPS模型组及LPS+RUB(25、50和100µmol/L)干预组,采用ELISA法检测细胞上清中白细胞介素1β(IL-1β)和肿瘤坏死因子α(TNF-α)的浓度;RTqPCR法检测促炎细胞因子环加氧酶2(COX-2)、诱导型一氧化氮合酶(iNOS)、IL-1β和TNF-α的mRNA表达水平;Western blot法检测NF-κB和MAPK信号通路相关蛋白的表达;免疫荧光染色观察细胞中p65的表达情况。结果:与对照组相比,LPS诱导后细胞上清液中IL-1β和TNF-α的分泌量增加(P<0.01),促炎细胞因子COX-2、iNOS、IL-1β和TNF-α的mRNA水平显著增加(P<0.01),NF-κB和MAPK信号通路蛋白表达显著上调(P<0.05或P<0.01),且p65出现荧光强度增加(P<0.05);与LPS模型组相比,LPS+RUB(25、50和100µmol/L)干预组中,IL-1β和TNF-α的分泌量减少(P<0.05或P<0.01),促炎细胞因子COX-2、iNOS、IL-1β和TNF-α的mRNA水平降低(P<0.05或P<0.01),p-p65、p-IκBα、p-ERK、p-p38和p-JNK蛋白表达显著下调(P<0.05或P<0.01),同时p65荧光减弱(P<0.05)。结论:体外实验表明RUB能够通过抑制NF-κB和MAPK信号通路的活化减轻小鼠神经炎症细胞模型炎症反应。AIM:The effect of rubusoside(RUB)on neuroinflammatory response of BV-2 microglia in mice induced by lipopolysaccharide(LPS)was investigated in vitro.METHODS:CCK-8 assay was used to detect the effect of RUB on cell viability.BV-2 cells were divided into control group,LPS model group and LPS+RUB(25,50 and 100µmol/L)intervention group.The concentrations of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in the supernatant of BV-2 microglia were detected by ELISA.RT-qPCR was used to detect the mRNA expression levels of proinflammatory cytokine cyclooxygenase-2(COX-2),inducible nitric oxide synthase(iNOS),IL-1βand TNF-α.The protein expression of NF-κB and MAPK pathway was detected by Western blot.The expression of p65 in cells was observed by immunofluorescence staining.RESULTS:Compared with control group,IL-1βand TNF-αin LPS-induced cell supernatant(P<0.01),the mRNA levels of proinflammatory cytokines COX-2,iNOS,IL-1βand TNF-αwere significantly increased(P<·616·0.01),NF-κB and MAPK signaling pathway proteins were significantly up-regulated(P<0.05 or P<0.01)and increased fluorescence intensity in p65(P<0.05).Compared with the LPS model group,the levels of IL-1β(P<0.01)and TNF-α(P<0.05),the mRNA levels of proinflammatory cytokines COX-2,iNOS,IL-1βand TNF-αwere decreased(P<0.05 or P<0.01),the protein expressions of p-p65,p-IκBα,p-ERK,p-p38 and p-JNK were significantly down-regulated(P<0.05 or P<0.01),while p65 fluorescence decreased(P<0.05).CONCLUSION:In vitro experiments have shown that RUB can reduce the inflammatory response of mouse neuroinflammatory cell models by inhibiting the activation of NF-κB and MAPK signaling pathways.

关 键 词：甜叶悬钩子苷 神经炎症 小胶质细胞 NF-ΚB信号通路 MAPK信号通路 

分 类 号：R745.1[医药卫生—神经病学与精神病学] R363.2[医药卫生—临床医学]