miR-322 treatment rescues cell apoptosis and neural tube defect formation through silencing NADPH oxidase 4
miR-322 treatment rescues cell apoptosis and neural tube defect formation through silencing NADPH oxidase 4
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miR-322 治疗通过沉默 NADPH 氧化酶 4 来挽救细胞凋亡和神经管缺陷形成
DOI：
10.1111/cns.13383
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发表时间：
2020-04-24
影响因子：
5.5
通讯作者：
Yuan, Zheng-wei
中科院分区：
文献类型：
--
作者：
Liu, Yu-si;Gu, Hui;Yuan, Zheng-wei
目的神经细胞凋亡过多导致神经管闭合失败导致神经管缺损。NADPH氧化酶4 （NOX4）是细胞生长和死亡的关键介质，但其在NTDs中的作用从未被表征。NOX4是miR-322的潜在靶点，我们之前已经证明miR-322参与高糖诱导的NTDs。在这项研究中，我们研究了NOX4对NTDs胚胎神经上皮的影响，并揭示了miR-322通过改善细胞凋亡来破坏神经调节的新机制。方法采用全反式维甲酸（ATRA）诱导小鼠ntd模型。采用RNA下拉和双荧光素酶报告基因检测来证实NOX4和miR-322之间的相互作用。在小鼠神经干细胞和全胚胎培养中，采用Western blot和TUNEL方法研究miR-322和NOX4对NTD形成过程中神经上皮细胞凋亡的影响。结果NOX4作为miR-322的新靶点，在atra诱导的NTDs小鼠模型中表达上调。在小鼠神经干细胞中，miR-322可抑制NOX4的表达；进一步，nox4触发的细胞凋亡也被miR-322抑制。此外，在全胚胎培养中，将miR-322模拟物注射到羊膜腔中，通过沉默NOX4减少NTD形成过程中的细胞凋亡。结论miR-322/NOX4在细胞凋亡诱导的NTD形成中起着至关重要的作用，这可能为胚胎NTDs的机制提供新的认识，并为寻找潜在的治疗靶点提供依据。
Aims Failure of neural tube closure resulting from excessive apoptosis leads to neural tube defects (NTDs). NADPH oxidase 4 (NOX4) is a critical mediator of cell growth and death, yet its role in NTDs has never been characterized. NOX4 is a potential target of miR-322, and we have previously demonstrated that miR-322 was involved in high glucose-induced NTDs. In this study, we investigated the effect of NOX4 on the embryonic neuroepithelium in NTDs and reveal a new regulatory mechanism for miR-322 that disrupts neurulation by ameliorating cell apoptosis.Methods All-trans-retinoic acid (ATRA)-induced mouse model was utilized to study NTDs. RNA pull-down and dual-luciferase reporter assays were used to confirm the interaction between NOX4 and miR-322. In mouse neural stem cells and whole-embryo culture, Western blot and TUNEL were carried out to investigate the effects of miR-322 and NOX4 on neuroepithelium apoptosis in NTD formation.Results NOX4, as a novel target of miR-322, was upregulated in ATRA-induced mouse model of NTDs. In mouse neural stem cells, the expression of NOX4 was inhibited by miR-322; still further, NOX4-triggered apoptosis was also suppressed by miR-322. Moreover, in whole-embryo culture, injection of the miR-322 mimic into the amniotic cavity attenuated cell apoptosis in NTD formation by silencing NOX4.Conclusion miR-322/NOX4 plays a crucial role in apoptosis-induced NTD formation, which may provide a new understanding of the mechanism of embryonic NTDs and a basis for potential therapeutic target against NTDs.