白藜芦醇减轻同型半胱氨酸引起的巨噬细胞免疫应答和增殖的作用  

Resveratrol alleviates the immune response and proliferation of macrophages induced by homocysteine

作　　者：张学森 秦凯悦 李晓菡 王艳佳 徐茜 杨晓玲 ZHANG Xue-sen;QIN Kai-yue;LI Xiao-han;WANG Yan-jia;XU Xi;YANG Xiao-ling(Department of Orthopaedics,People's Hospital of Wuzhong,Wuzhong 751199,Ningxia Hui Autonomous Region,China;Key Laboratory of Metabolic Cardiovascular Disease Research of National Board of Health,Yinchuan 750004,Ningxia Hui Autonomous Region,China;Department of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Hui AutonomousR egion,China)

机构地区：[1]吴忠市人民医院骨科,宁夏回族自治区吴忠751199 [2]国家卫健委代谢性心血管疾病研究重点实验室,宁夏回族自治区银川750004 [3]宁夏医科大学基础医学院,宁夏回族自治区银川750004

出　　处：《中国临床药理学杂志》2024年第14期2038-2042,共5页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81960063);宁夏自然科学基金资助项目(2022AAC03765,2021AAC02012)。

摘　　要：目的探讨白藜芦醇(Res)在同型半胱氨酸(Hcy)引起巨噬细胞免疫应答和增殖中的作用。方法将小鼠ANA-1细胞分为对照组(常规培养)、模型组(100μmol·L^(-1) Hcy)和低、中、高剂量实验组(在模型组基础上分别加25、50和100μmol·L^(-1)白藜芦醇)以及Hcy+Ad-SIRT1组(100μmol·L^(-1) Hcy+Ad-SIRT1)、Hcy+si-FOXO1组(100μmol·L^(-1) Hcy+si-FOXO1)、Hcy+Res-L+Ad-SIRT1+si-FOXO1组(100μmol·L^(-1) Hcy+25μmol·L^(-1)白藜芦醇+转染Ad-SIRT1+si-FOXO1)。用四甲基偶氮唑蓝(MTT)法检测细胞增殖,用酶联免疫吸附法检测细胞培养液上清液中白细胞介素-6(IL-6)和肿瘤坏死因子-α(TNF-α)的浓度。用蛋白质印迹法检测沉默信息调节因子(SIRT1)和叉头蛋白O1(FOXO1)蛋白的表达水平。结果对照组、模型组和低、中、高剂量实验组450 nm光密度值分别为0.25±0.02、0.36±0.02、0.33±0.01、0.30±0.02和0.29±0.01,模型组较对照组细胞增殖显著增加,低、中、高剂量实验组细胞增殖较模型组均显著减少(均P<0.05);对照组、模型组、低剂量实验组细胞培养液上清液中IL-6分别为(394.04±20.06)、(614.23±21.09)和(501.53±16.52)pg·mL^(-1),TNF-α分别为(516.54±18.96)、(717.22±24.81)和(632.74±19.11)pg·mL^(-1),SIRT1蛋白相对表达水平分别1.00±0.05、0.57±0.05和0.77±0.04,FOXO1蛋白相对表达水平分别为1.00±0.05、2.31±0.18和1.58±0.11,低剂量实验组的上述指标与对照组比较,在统计学上差异均有统计学意义(均P<0.05)。模型组、低剂量实验组、Hcy+Ad-SIRT1组及Hcy+si-FOXO1组细胞培养液上清液IL-6和TNF-α含量均显著低于模型组,在统计学上差异有统计学意义(均P<0.05);低剂量实验组共转染Ad-SIRT1和si-FOXO1后,细胞培养液上清液IL-6和TNF-α含量均显著低于Ad-SIRT1组和si-FOXO1组(均P<0.05)。结论白藜芦醇能够减轻Hcy引起的巨噬细胞免疫应答和增殖,其机制与SIRT1/FOXO1通路有关。Objective To explore the role of resveratrol in the immune response and proliferation of macrophages induced by homocysteine(Hcy).Methods ANA-1 cells were divided into control group(conventional culture),model group(100μmol·L^(-1)Hcy),experimental-L,-M,-H groups(adding 25,50 and 100μmol·L^(-1)resveratrol to model group,respectively),Hcy+Ad-SIRT1 group(100μmol·L^(-1)Hcy+Ad-SIRT1),Hcy+si-FOXO1 group(100μmol·L^(-1)Hcy+si-FOXO1),Hcy+Res-L+Ad-SIRT1+si-FOXO1 group(100μmol·L^(-1)Hcy+25μmol·L^(-1)Resveratrol transfected with Ad-SIRT1+si-FOXO1).The cell proliferation was detected by methyl thiazolyl tetrazolium(MTT),and the concentration of interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in the supernatant of cell culture medium was detected by enzyme-linked immunosorbent assay.The gene and protein expression of silencing information regulator 1(SIRT1)and forkhead protein O1(FOXO1)were detected by Western blot.Results The optical density of 450 nm in control group,model group and experimental-L,-M,-H groups were 0.25±0.02,0.36±0.02,0.33±0.01,0.30±0.02 and 0.29±0.01,respectively.Compared with the control group,the cell proliferation in the model group was significantly increased(P<0.05).Cell proliferation in experimental-L,-M,-H groups was significantly decreased compared with model group(all P<0.05).IL-6 in the supernatant of cell culture medium of control group,model group and experimental-L group were(394.04±20.06),(614.23±21.09)and(501.53±16.52)pg·mL^(-1),respectively;TNF-αwere(516.54±18.96),(717.22±24.81)and(632.74±19.11)pg·mL^(-1),respectively;SIRT1 relative protein expression were(1.00±0.05),(0.57±0.05)and(0.77±0.04),respectively;the relative protein expression of FOXO1 were1.00±0.05,2.31±0.18 and 1.58±0.11,respectively.Compared with the control group,the above indexes in the experimental-L group had statistical significance(all P<0.05).The contents of IL-6 and TNF-αin cell culture fluid supernatant in model group,experimental-L group,Hcy+Ad-SIRT1 group and Hcy+si-FOXO1 group

关 键 词：白藜芦醇 同型半胱氨酸 沉默信息调节因子1 叉头蛋白O1 免疫应答 增殖 

分 类 号：R28[医药卫生—中药学]