基于Super-SILAC定量技术系统解析槲皮素调控巨噬细胞的组蛋白翻译后修饰图谱  

Systematic Analysis of Histone Post-translational Modifications in Macrophages Regulated by Quercetin Based on Super-SILAC

作　　者：刘志晴 翟琳辉 吉敬 谭敏佳 刘萍 LIU Zhi-Qing;ZHAI Lin-Hui;JI Jing;TAN Min-Jia;LIU Ping(Jiangsu Key Laboratory of Marine Pharmaceutical Compound Screening,School of Pharmacy,Jiangsu Ocean University,Jiangsu,Lianyungang 222005,China;Shanghai Institute of Materia Medica,Chinese Academy of Sciences,Shanghai 201203,China;School of Chinese Materia Medica,Nanjing University Of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]江苏海洋大学药学院,江苏省海洋药物化合物筛选重点实验室,江苏连云港222005 [2]中国科学院上海药物研究所,上海201203 [3]南京中医药大学新中药学院,南京210023

出　　处：《中国生物化学与分子生物学报》2024年第11期1574-1584,共11页Chinese Journal of Biochemistry and Molecular Biology

基　　金：上海市优秀学术/技术带头人(No.22XD1420900)资助。

摘　　要：槲皮素(quercetin,Que)是一种具有多种生物活性的黄酮类化合物,在自然界中分布广泛,具有抗炎和抗肿瘤等生物活性。研究发现,槲皮素的抗炎活性与其对组蛋白翻译后修饰的调控密切相关。然而,槲皮素调控组蛋白翻译后修饰发挥抗炎作用的精细机制尚未有详细报道。为了探讨槲皮素调控组蛋白翻译后修饰发挥抗炎活性的作用机制,我们首先评估了槲皮素对M1型巨噬细胞极化的影响,结果发现,槲皮素可以明显下调M1型巨噬细胞的白细胞介素-1β(interleukin-1β,IL-1β)和白细胞介素-6(interleukin-6,IL-6)水平;其次,利用生物质谱的细胞稳定同位素标记(stable isotope labeling by amino acids in cell culture,SILAC)技术衍生的super-SILAC方法,进行了组蛋白翻译后修饰的解析,系统定量分析了槲皮素处理后的巨噬细胞中组蛋白修饰水平的全局性变化;最终,共定量到30个组蛋白修饰位点,其中包括12个发生下调的组蛋白乙酰化修饰位点和4个发生上调的甲基化修饰位点(fold change>1.2,P<0.05);进一步对部分差异变化位点进行了蛋白质免疫印迹(western blot,WB)验证,其差异变化与质谱结果一致性良好。综上,本研究系统解析了槲皮素调控巨噬细胞的组蛋白翻译后修饰图谱,为槲皮素抗炎作用机制的研究提供了可靠的数据参考和新的线索。Quercetin(Que)is a flavonoid compound widely distributed in nature with various biological activities.Its anti-inflammatory effect plays a crucial role in many diseases,closely related to its regulation of histone post-translational modifications.However,there have been no detailed reports on the anti-inflammatory effects of quercetin regulating histone post-translational modifications.In this study,we first investigated the effect of quercetin on the M1 macrophages polarization.The results showed that quercetin can significantly down-regulate the levels of interleukin-1β(IL-1β)and interleukin-6(IL-6)in M1 macrophages.Next,we used the super stable isotope labeling by amino acids in cell culture(super-SILAC)method derived from SILAC technology based on mass spectrometry to systematically analyze the post-translational modification levels of histone in macrophages treated with quercetin.A total of 30 histone modification sites were quantified,of which 12 histone lysine acetylation marks were significantly downregulated and 4 lysine methylation sites were upregulated(fold change>1.2,P<0.05),and some sites were verified by Western blot(WB),which was consistent with the mass spectrometry results.In conclusion,a comprehensive analysis of quercetin on regulating macrophage histone modifications in this study provides reliable data references and new insights for studying its anti-inflammatory mechanism.

关 键 词：槲皮素 表观遗传 组蛋白翻译后修饰 细胞稳定同位素标记 巨噬细胞 

分 类 号：Q946.885[生物学—植物学]