白洁,孟军,刘艳辉,李小红,危当恒.Ox-LDL对PMA诱导的THP-1巨噬细胞自噬的影响.[J].中南医学科学杂志.,2014,42(3):217-221.
Ox-LDL对PMA诱导的THP-1巨噬细胞自噬的影响
The Effect of Ox-LDL on Autophagy in PMA InducedTHP-1-derived Macrophages
投稿时间:2014-01-02  
DOI:
中文关键词:  Ox-LDL  THP-1巨噬细胞  自噬
英文关键词:oxidized low density lipoprotein  THP-1 macrophage  autophagy
基金项目:国家自然科学基金资助项目(81370378).
作者单位
白洁1,孟军1,刘艳辉2,李小红2,危当恒2 1.南华大学附属第一医院功能科2.南华大学心血管疾病研究所动脉硬化学湖南省重点实验室湖南 衡阳 421001 
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中文摘要:
      目的探讨氧化型低密度脂蛋白(Ox-LDL)对PMA诱导的THP-1巨噬细胞自噬的影响。方法PMA诱导THP-1细胞24 h,使其分化为巨噬细胞,再分别用0、10、20、40或80 mg/L的Ox-LDL处理24 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹技术(Western blotting)分别检测Beclin-1以及LC3的mRNA及蛋白表达;细胞免疫荧光检测LC3在细胞内的含量;MDC染色检测自噬囊泡的形成。结果随着Ox-LDL浓度的增加,THP-1源性巨噬细胞Beclin-1 mRNA及蛋白表达显著降低(P<0.05);LC3 mRNA表达无明显改变(P>0.05),但蛋白水平LC3II/LC3I显著降低(P<0.001);免疫荧光结果表明随着Ox-LDL浓度的增加,LC3II含量降低,MDC染色结果显示自噬囊泡随着Ox-LDL浓度的增加而减少。结论Ox-LDL抑制PMA诱导的THP-1巨噬细胞自噬。
英文摘要:
      ObjectiveTo explore the effect of oxidized low density lipoprotein (Ox-LDL) on autophagy in THP-1-derived macrophages induced by phorbol-12-myristate-13-acetate (PMA).MethodsAfter treatment with PMA for 24 h,THP-1-derived macrophages were treated with 0,10,20,40 or 80 mg/L Ox-LDL for 24 h.Reverse transcription polymerase chain reaction (RT-PCR) and Western blotting were used to detect Beclin 1 and LC3 mRNA and protein expression.Cell immunofluorescence was used to detect intracellular LC3II protein concentration.The autofluorescent substance monodansylcadaverine (MDC) was used to detect autophagic vacuoles (AVs).ResultsAfter treated with 0,10,20,40 or 80 mg/L Ox-LDL for 24 h,Beclin-1 mRNA and protein expression were obviously decreased(P<0.05).There was no obvious change in LC3 mRNA expression (P>0.05).However,the ratio of LC3II/LC3I was decreased with a concentration dependent manner at the level of protein (P<0.001).Cell immunofluorescence showed that the intracellular LC3II concentration was decreased in response to Ox-LDL concentration.MDC staining showed that AVs had negative correlation with Ox-LDL concentration.ConclusionOx-LDL inhibited PMA induced-THP-1 macrophage autophagy.
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