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Role of catechins on ET-1-induced stimulation of PLD and NADPH oxidase activities in pulmonary smooth muscle cells: determination of the probable mechanism by molecular docking studies

机译:儿茶素对肺部平滑肌细胞中的ET-1诱导刺激PLD和NADPH氧化酶活性的作用:通过分子对接研究确定可能的机制

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摘要

The treatment of human pulmonary artery smooth muscle cells with ET-1 stimulates the activity of PLD and NADPH oxidase, but this stimulation is inhibited by pretreatment with bosentan (ET-1 receptor antagonist), FIPI (PLD inhibitor), apocynin (NADPH oxidase inhibitor), and EGCG and ECG (catechins having a galloyl group), but not EGC and EC (catechins devoid of a galloyl group). Herein, using molecular docking analyses based on our biochemical studies, we determined the probable mechanism by which the catechins containing a galloyl group inhibit the stimulation of PLD activity induced by ET-1. The ET-1-induced stimulation of PLD activity was inhibited by SecinH3 (inhibitor of cytohesin). Arf6 and cytohesin-1 are associated in the cell membrane, which is not inhibited by the catechins during ET-1 treatment of the cells. However, EGCG and ECG inhibited the binding of GTP gamma S with Arf6, even in the presence of cytohesin-1. The molecular docking analyses revealed that the catechins containing a galloyl group (EGCG and ECG) with cytoliesin-l-Arf6(GDP), but not the catechins without a galloyl group (EGC and EC), prevent GDP-GTP exchange in Arf6, which seems to be an important mechanism for inhibiting the activation of PLD induced by ET-1, and subsequently increases the activity of NADPH oxidase.
机译:用ET-1治疗人肺动脉平滑肌细胞刺激PLD和NADPH氧化酶的活性,但是通过与波丝坦(ET-1受体拮抗剂),FIPI(PLD抑制剂),鼻咽(NADPH氧化酶抑制剂)进行预处理来抑制这种刺激。(NADPH氧化酶抑制剂)和EGCG和ECG(具有储华基团的儿茶素),但不是EGC和EC(儿茶素缺乏Galloyl组)。这里,使用基于我们的生化研究的分子对接分析,我们确定了含Galloyl基团的儿茶素抑制ET-1诱导的PLD活性刺激的可能机制。 SecinH3(细胞素抑制剂)抑制了ET-1诱导的PLD活性刺激。 ARF6和细胞蛋白-1在细胞膜中涉及,其在细胞的ET-1治疗期间不受儿茶素抑制。然而,即使在细胞蛋白-1存在下,EGCG和ECG也抑制了GTPγs与ARF6的结合。分子对接分析表明,含有细胞蛋白酶-1-ARF6(GDP)的加入酰基(EGCG和ECG),但不是没有Galloyl基团(EGC和EC)的儿茶素,防止ARF6中的GDP-GTP交换,这似乎是抑制ET-1诱导的PLD活化的重要机制,随后增加了NADPH氧化酶的活性。

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