摘要
目的探讨白藜芦醇(resveratrol,RES)抑制白介素1β(interleukin 1β,IL-1β)处理的SW1353细胞的炎症反应与叉头框转录因子O1(forkhead transcription factors O1,FoxO1)/细胞沉默信息调节子1(silent information regulator 1,Sirt1)信号之间的关系。方法采用SW1353软骨肉瘤细胞系,进行如下4个方面的处理及检测:给予10ng/ml IL-1β的同时,加入RES(12.5、50μmol/L)处理24 h,Western blot法检测Sirt1蛋白的表达;给予10ng/mL IL-1β的同时,加入RES(50μmol/L)处理30 min,免疫细胞化学技术检测FoxO1的定位,Western blot法检测FoxO1及p-FoxO1水平;FoxO1 siRNA进行转染,荧光显微镜下观察转染效率,Western blot法检测FoxO1的抑制效果及对Sirt1的影响;FoxO1siRNA转染后,Western blot法检测RES处理对Sirt1蛋白表达的影响,ELISA法检测细胞培养基上清IL-6的水平。结果单纯IL-1β处理对Sirt1的蛋白表达无明显影响,而给予RES(12.5、50μmol/L)处理后显著增加Sirt1水平。加入IL-1β/RES后,FoxO1主要定位于细胞质中;IL-1β处理显著上调p-FoxO1水平,而加入RES则进一步增加p-FoxO1水平;FoxO1水平保持不变。FoxO1基因沉默可有效抑制细胞FoxO1水平,同时Sirt1水平也被显著抑制。FoxO1基因沉默后抑制RES诱导的Sirt1水平,抑制IL-1β增加的IL-6水平,而RES处理则引起IL-6水平的进一步降低。结论在IL-1β处理的SW1353细胞中,RES可以通过失活FoxO1,增加Sirt1的水平发挥抗炎效应。
Objective To explore whether the anti-osteoarthritis(OA)effect of resveratrol(RES)on IL-1β-induced SW1353 cells is related to forkhead transcription factors O1(FoxO1)and silent information regulator 1(Sirt1).Methods SW1353 chondrosarcoma cell line was treated with the following methods:IL-1β(10ng/ml)and RES(12.5,50μmol/L)co-treated for 24 h,and then Sirt1 protein was detected by Western blot;IL-1β(10ng/ml)and RES(50μmol/L)co-treated for 30 min,and then the localization of FoxO1 was detected by immunocytochemistry,and the protein expression of FoxO1 and p-FoxO1 was detected by western blot;FoxO1 siRNA was transfected and the transfection efficiency was observed under fluorescence microscope,then the inhibitory effect of FoxO1 and the effect on Sirt1 were detected by western blot;after FoxO1 siRNA transfection,the effect of RES on Sirt1 protein expression was detected by western blot,and the level of IL-6 in cell culture supernatant was detected by ELISA.Results IL-1βtreatment had no significant effect on Sirt1 protein expression,while RES(12.5,50μmol/L)treatment significantly increased Sirt1 protein expression.FoxO1 was mainly localized in cytoplasm after IL-1βand/or RES treatment;IL-1βtreatment significantly up-regulated p-FoxO1 level,and addition of RES further increased p-FoxO1 expression;however,FoxO1 expression remained unchanged.FoxO1 gene silencing could effectively inhibit the expression of FoxO1,as well as the protein expression of Sirt1.FoxO1 gene silencing inhibited RES-in duced Sirt1 protein expression;and inhibited IL-1β-upregulated IL-6 level,while RES treatment caused a further decrease in IL-6 level.Conclusion In IL-1β-treated SW1353 cells,RES can exert an anti-inflammatory effect by inactivating FoxO1 and increasing the level of Sirt1.
作者
顾海伦
徐小磊
刘旭丹
刘莉
Gu Hailun;Xu Xiaolei;Liu Xudan;Liu Li(Department of Orthopedics,Shengjing Hospital,China Medical University;Department of Nutrition and Food Hygiene,School of Public Health,China Medical University,Shenyang 110004,China)
出处
《中国组织化学与细胞化学杂志》
CAS
CSCD
2019年第4期311-317,共7页
Chinese Journal of Histochemistry and Cytochemistry
基金
国家自然科学基金(81372971)
沈阳市科技计划项目(RC170476)
