Effect of Oxidized Low Density Lipoprotein on TET2 Expression and Autophagy of Human Umbilical Vein Endothelial Cell
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摘要:
目的观察氧化型低密度脂蛋白对人脐静脉内皮细胞自噬的影响及其调节机制。方法氧化型低密度脂蛋白孵育人脐静脉内皮细胞 24 h, RT-PCR检测TET2 mRNA表达,Western blot 检测TET2以及自噬标记物Beclin 1、LC3的表达。TET2 siRNA转染人脐静脉内皮细胞,Western blot 检测Beclin 1、LC3的表达。结果人脐静脉内皮细胞经不同浓度氧化型低密度脂蛋白孵育24 h后,浓度依赖性地下调TET2 mRNA 以及蛋白的表达(p<0.05),并且下调Beclin 1表达以及LC3Ⅱ/LC3Ⅰ的比值。TET2 siRNA 转染人脐静脉内皮细胞后,Beclin 1表达以及LC3Ⅱ/LC3Ⅰ的比值明显降低。结论氧化型低密度脂蛋白促进人脐静脉内皮细胞自噬,可能与下调TET2表达相关。
Abstract:
AimTo investigate the effect of oxidized low density lipoprotein (ox-LDL) on ten-eleven-translocation 2 (TET2) expression and autophagy in human umbilical vein endothelial cells (HUVEC).MethodsAfter treatment with ox-LDL for 24 h, RT-PCR was used to detect mRNA expression of TET2.Western blot was used to detect protein expression of TET2 and autophagic marker Beclin 1 and microtubule-associated protein light chain 3 (LC3).HUVEC were transfected with TET2 siRNA, and then Beclin 1 and LC3 were detected by using Western blot.ResultsAfter treatment with ox-LDL for 24 h, the expression of TET2 was decreased with a concentration-depended manner both in mRNA and protein levels.The protein expression of Beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰ were increased with response to ox-LDL concentration.Transfection with TET2 siRNA, the expression of Beclin 1 and the ratio of LC3Ⅱ/LC3Ⅰ were upregulated.Conclusionox-LDL decreased the expression of TET2, which might contribute to the increase of HUVEC autophagy.