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作 者:刘亚娴 李剑勇[2] 李世宏[2] 杨亚军[2] 刘希望[2] 葛闻博 白莉霞 秦哲[2] 李存 LIU Yaxian;LI Jianyong;LI Shihong;YANG Yajun;LIU Xiwang;GE Wenbo;BAI Lixia;QIN Zhe;LI Cun(Tianjin Key Laboratory of Agricultural Animal Breeding and Healthy Husbandry,College of Animal Science and Veterinary Medicine/Tianjin Agricultural University,Tianjin 300392,China;Key Laboratory of New Animal Drug Project of Gansu Province,Key Laboratory of Veterinary Pharmaceutical Development of Ministry of Agriculture and Rural Affairs,Lanzhou Institute of Husbandry and Pharmaceutical Science of Chinese Academy Agriculture Sciences,Lanzhou 730050,China)
机构地区:[1]天津农学院动物科学与动物医学学院/天津市农业动物繁育与健康养殖重点实验室,天津300392 [2]中国农业科学院兰州畜牧与兽药研究所/农业农村部兽用药物创制重点实验室甘肃省新兽药工程重点实验室,兰州730050
出 处:《黑龙江畜牧兽医》2024年第3期16-21,30,共7页Heilongjiang Animal Science And veterinary Medicine
基 金:国家自然科学基金青年科学基金项目(32002341)。
摘 要:为了探讨阿司匹林丁香酚酯(AEE)对氧化型低密度脂蛋白(ox-LDL)诱导的RAW264.7细胞中诱导型一氧化氮合酶(iNOS)表达及一氧化氮(NO)合成的影响,试验首先采用CCK-8法检测细胞存活率,采用油红O染色法观察细胞内脂滴形成情况,采用实时荧光定量PCR扩增、免疫荧光技术和Western-blot技术测定iNOS mRNA和蛋白的相对表达量,最后通过硝酸盐还原酶测定法测定NO含量。结果表明:AEE和ox-LDL对RAW264.7细胞没有毒性。利用80μg/mL的ox-LDL可成功诱导RAW264.7细胞泡沫化,建立泡沫细胞模型,并且AEE可明显抑制细胞内脂滴的形成。ox-LDL可极显著上调细胞中iNOS mRNA及蛋白的相对表达量(P<0.01),并且极显著增加了细胞内NO含量(P<0.01)。经AEE预处理的RAW264.7细胞中iNOS mRNA及蛋白相对表达量显著或极显著降低(P<0.05或P<0.01),细胞内NO含量也极显著减少(P<0.01)。说明AEE可通过抑制iNOS的表达降低NO的合成,缓解RAW264.7细胞泡沫化,进而发挥抗动脉粥样硬化的作用。To investigate the effects of Aspirin eugenol ester(AEE) on inducible nitric oxide synthase(iNOS) expression and nitric oxide(NO) synthesis in RAW264.7 cells induced by oxidized low density lipoprotein(ox-LDL),the cell survival rate was detected by CCK-8 method in the experiment.Oil red O staining was used to observe the formation of intracellular lipid droplets.The relative expression of iNOS mRNA and protein was detected by real-time fluorescent quantitative PCR,immunofluorescence and Western-blot.Finally,the content of NO was determined by nitrate reductase assay.The results showed that AEE and ox-LDL were not toxic to RAW264.7 cells.RAW264.7 macrophages were successfully induced to foam using 80 μg/mL of ox-LDL to establish a foam cell model,and AEE significantly inhibited the formation of intracellular lipid droplets.ox-LDL significantly upregulated the expression of iNOS mRNA and protein in the cells(P<0.01) and significantly increased the intracellular NO content(P<0.01).The expression of iNOS mRNA and protein in RAW264.7 cells pretreated with AEE was significantly decreased(P<0.05 or P<0.01),and the content of NO in RAW264.7 cells was also significantly decreased(P<0.01).These results indicated that AEE could inhibit the expression of iNOS and reduce the synthesis of NO,alleviate the formation of RAW264.7 foam cells,and play a role in anti-atherosclerosis.
关 键 词:动脉粥样硬化 RAW264.7 泡沫细胞 一氧化氮 诱导型一氧化氮合酶
分 类 号:S859.7[农业科学—临床兽医学]
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