ox-LDL经由LOX-1受体对PPARγ-LXRα-ABCA1通路激活的作用研究
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(1.辽宁省糖脂代谢重点实验室 大连大学生命科学与技术学院,辽宁省大连市 116600;2.大连理工大学生命科学与技术学院,辽宁省大连市 116024)

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余承洁,硕士研究生,研究方向为动脉粥样硬化细胞分子生物学,E-mail为13478916083@163.com。

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国家自然科学基金项目(81270361)


Ox-LDL activate PPARγ-LXRα-ABCA1 to transcript by LOX-1 receptor
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1.Key Laboratory of Glucolipid Metabolism in Liaoning Province & College of Life Science and Technology, Dalian University, Dalian, Liaoning 116600, China;2.College of Life Science and Technology, Dalian University of Technology, Dalian, Liaoning 116024, China)

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    摘要:

    目的 探讨氧化型低密度脂蛋白(ox-LDL)经由血凝素样氧化型低密度脂蛋白受体1(LOX-1)受体对PPARγ-LXRα-ABCA1通路激活的作用。 方法 浓度梯度ox-LDL(0~40 mg/L,12 h)刺激J774A.1巨噬细胞,检测LXRα、ABCA1的表达。构建293T细胞LOX-1过表达的PPARγ双荧光素酶报告基因系统,激光共聚焦及Western blot检测LOX-1的分布与表达,双荧光素酶报告基因系统检测PPARγ的转录活化情况。对J774A.1巨噬细胞分别进行LOX-1 siRNA和PPARγ siRNA沉默,ox-LDL(30 mg/L,12 h)孵育后检测LXRα、ABCA1蛋白的表达。 结果 ox-LDL可显著上调J774A.1细胞LXRα和ABCA1的表达(P<0.01,n=3)。LOX-1过表达的PPARγ双荧光素酶报告基因系统检测显示ox-LDL能够通过LOX-1增加PPARγ的转录活性;对J774A.1巨噬细胞分别进行LOX-1 siRNA、PPARγ siRNA沉默后发现,LXRα和ABCA1的表达均显著降低(P<0.01,n=3)。 结论 ox-LDL可通过LOX-1受体激活PPARγ转录活性,从而上调LXRα、ABCA1蛋白表达,完成PPARγ-LXRα-ABCA1信号通路的激活。

    Abstract:

    Aim To investigate the effect of oxidized low density lipoprotein (ox-LDL) on activation of PPARγ-LXRα-ABCA1 pathway via lectin-like oxidized low density lipoprotein receptor-1 (LOX-1) receptor. Methods The expression of liver X receptor α (LXRα) and ABCA1 was detected by stimulating J774A.1 macrophages with concentration gradient ox-LDL (0~40 mg/L, 12 h), the expression of LOX-1 was detected by laser confocal microscopy and Western blot, the transcriptional activation of peroxisome proliferators-activated receptor γ (PPARγ) was detected by double luciferase reporter gene, the expression of LXRα and ATP-binding cassette transporter A1 (ABCA1) protein was detected after incubation with J774A.1 macrophages by LOX-1 siRNA and PPARγ siRNA silencing, ox-LDL (30 mg/L,12 h). Results Ox-LDL significantly up-regulated the expression of LXRα and ABCA1 in J774A.1 cells (significantly different from the cell untreated by ox-LDL as control, P<0.01, n=3). LOX-1 overexpression of PPARγ double luciferase reporter gene system showed that ox-LDL increased the transcriptional activity of PPARγ by LOX-1. LOX-1 siRNA and PPARγ siRNA were treated with J774A.1 macrophages, respectively, and LXRα and ABCA1 were expressed (significantly different from the cell untreated by siRNA as control, P<0.01, n=3). Conclusion Ox-LDL activates PPARγ transcriptional activity by LOX-1 receptor, thereby upregulating LXRα and ABCA1 protein expression and activating PPARγ-LXRα-ABCA1 signal pathway.

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余承洁,李敬达,修志龙,刘庆平. ox-LDL经由LOX-1受体对PPARγ-LXRα-ABCA1通路激活的作用研究[J].中国动脉硬化杂志,2017,25(10):978~984.

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  • 收稿日期:2017-02-07
  • 最后修改日期:2017-03-20
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  • 在线发布日期: 2017-11-28