NF-κB-Gasdermin D (GSDMD) Axis Couples Oxidative Stress and NACHT, LRR and PYD Domains-Containing Protein 3 (NLRP3) Inflammasome-Mediated Cardiomyocyte Pyroptosis Following Myocardial Infarction.
NF-κB-Gasdermin D (GSDMD) Axis Couples Oxidative Stress and NACHT, LRR and PYD Domains-Containing Protein 3 (NLRP3) Inflammasome-Mediated Cardiomyocyte Pyroptosis Following Myocardial Infarction.
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NF-κB-Gasdermin D（GSDMD）轴偶联氧化应激，NACHT，LRR和PYD结构域含有含量的蛋白3（NLRP3）炎症体炎症体介导的心肌细胞性心肌细胞凋亡后，心肌梗死后。
DOI：
10.12659/msm.908529
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发表时间：
2018-08-30
期刊：
影响因子：
--
通讯作者：
Chen C
中科院分区：
文献类型：
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作者：
Lei Q;Yi T;Chen C
下垂和氧化应激在心肌梗死后心肌细胞丢失中起关键作用。核因子-κB与氧化应激和胃泌素D(GSDMD)有关，后者是上睑下垂的效应分子。然而，氧化应激与心肌细胞下垂之间的确切关系仍不清楚。我们通过膜孔形成、乳酸脱氢酶(LDH)的释放以及caspase-1、裂解的caspase-1、Nacht、LRR和PYD结构域包含蛋白3(NLRP3)以及含有卡片的凋亡相关的SPECK样蛋白(ASC)的表达来检测炎症小体介导的在体心肌细胞松弛。此外，我们还在体外通过缺氧缺糖(OGD)诱导H9C2细胞发生热下垂。乳酸脱氢酶试剂盒和Western印迹检测证实NLRP3炎性小体介导的下垂。以活性氧(ROS)和超氧化物歧化酶(SOD)活性为指标评价氧化应激。我们用N-乙酰-半胱氨酸抑制氧化应激，并通过乳酸脱氢酶试剂盒和Western印迹检测NF-κ、B-GSDMD轴和下垂的变化。然后，用吡咯烷二硫代氨基甲酸酯抑制NF-κB的激活，并用定量逆转录聚合酶链式反应、Western印迹和乳酸脱氢酶检测试剂盒检测GSDMD活性和下垂的变化。NAC抑制氧化应激可降低H9C2细胞中NF-κB和GSDMD的活性，增加H9C2细胞的下垂，表现为乳酸脱氢酶的释放和NLRP3炎性小体的激活。此外，抑制NF-κB的激活可降低缺氧状态下H9C2细胞的GSDMD转录和激活以及NLRP3炎性小体介导的下垂。我们证明了NF-κB-GSDMD轴在氧化应激和NLRP3炎性小体介导的心肌细胞松弛之间起着桥梁作用。我们的发现为心肌梗死相关的心室重构的机制提供了重要的洞察力。
Pyroptosis and oxidative stress play pivotal roles in cardiomyocyte loss after myocardial infarction. NF-κB is associated with oxidative stress and gasdermin D (GSDMD), the effector molecule of pyroptosis. However, the exact relationship between oxidative stress and cardiomyocyte pyroptosis remains unknown. We measured inflammasome-mediated cardiomyocyte pyroptosis in vivo via membrane pore formation, lactate dehydrogenase (LDH) release, and expression of caspase-1, cleaved caspase-1, NACHT, LRR and PYD domains-containing protein 3 (NLRP3), and apoptosis-associated speck-like protein containing a CARD (ASC). Furthermore, we induced pyroptosis in vitro by oxygen-glucose deprivation (OGD) in H9C2 cells. NLRP3 inflammasome-mediated pyroptosis was confirmed by LDH assay kit and Western blot. Oxidative stress was evaluated by reactive oxygen species (ROS) and superoxide dismutase (SOD) activity. We suppressed oxidative stress with N-acetyl-cysteine (NAC) and measured subsequent changes to the NF-κB-GSDMD axis and pyroptosis by LDH assay kit and Western blot. Then, we inhibited NF-κB activation with pyrrolidine dithiocarbamate (PDTC) and measured changes to GSDMD activity and pyroptosis by qRT-PCR, Western blot, and LDH assay kit. Suppression of oxidative stress by NAC reduced NF-κB and GSDMD activation and increased pyroptosis, characterized by LDH release and NLRP3 inflammasome activation in H9C2 cells under OGD. Moreover, inhibition of NF-κB activation reduced GSDMD transcription and activation and NLRP3 inflammasome-mediated pyroptosis of H9C2 cells under OGD. We demonstrated that the NF-κB-GSDMD axis functioned as a bridge between oxidative stress and NLRP3 inflammasome-mediated cardiomyocyte pyroptosis. Our findings provide important insight into the mechanism of myocardial infarction-related ventricular remodeling.