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Persulfidation of transcription factor FOXO1 at cysteine 457: A novel mechanism by which H2S inhibits vascular smooth muscle cell proliferation
Journal of Advanced Research ( IF 10.7 ) Pub Date : 2020-07-01 , DOI: 10.1016/j.jare.2020.06.023
Xiaoyu Tian 1, 2 , Dan Zhou 3 , Yong Zhang 3 , Yunjia Song 1 , Qingyou Zhang 1 , Dingfang Bu 1 , Yan Sun 1 , Liling Wu 4, 5 , Yuan Long 3 , Chaoshu Tang 4, 5 , Junbao Du 1, 4 , Yaqian Huang 1 , Hongfang Jin 1, 2
Affiliation  

Introduction

The proliferation of vascular smooth muscle cells (VSMCs) is an important physiological and pathological basis for many cardiovascular diseases. Endogenous hydrogen sulfide (H2S), the third gasotransmitter, is found to preserve vascular structure by inhibiting VSMC proliferation. However, the mechanism by which H2S suppresses VSMC proliferation has not been fully clear.

Objectives

This study aimed to explore whether H2S persulfidates the transcription factor FOXO1 to inhibit VSMC proliferation.

Methods

After the proliferation of VSMC A7r5 cells was induced by endothelin-1 (ET-1), FOXO1 phosphorylation and proliferating cell nuclear antigen (PCNA) expression were detected by Western blotting, the degree of FOXO1 nuclear exclusion and PCNA fluorescent signals in the nucleus were detected by immunofluorescence, and the persulfidation of FOXO1 was measured through a biotin switch assay.

Results

The results showed that ET-1 stimulation increased cell proliferation, FOXO1 phosphorylation and FOXO1 nuclear exclusion to the cytoplasm in the cells. However, pretreatment with NaHS, an H2S donor, successfully abolished the ET-1-induced increases in the VSMC proliferation, FOXO1 phosphorylation, and FOXO1 nuclear exclusion to the cytoplasm. Mechanistically, H2S persulfidated the FOXO1 protein in A7r5 and 293T cells, and the thiol reductant DTT reversed this effect. Furthermore, the C457S mutation of FOXO1 abolished the H2S-induced persulfidation of FOXO1 in the cells and the subsequent inhibitory effects on FOXO1 phosphorylation at Ser256, FOXO1 nuclear exclusion to the cytoplasm and cell proliferation.

Conclusion

Thus, our findings demonstrated that H2S might inhibit VSMC proliferation by persulfidating FOXO1 at Cys457 and subsequently preventing FOXO1 phosphorylation at Ser256.



中文翻译:

转录因子 FOXO1 在半胱氨酸 457 上的过硫化:H2S 抑制血管平滑肌细胞增殖的新机制

介绍

血管平滑肌细胞(VSMCs)的增殖是许多心血管疾病的重要生理和病理基础。内源性硫化氢 (H 2 S) 是第三种气体递质,被发现通过抑制 VSMC 增殖来保护血管结构。然而,H 2 S 抑制 VSMC 增殖的机制尚不完全清楚。

目标

本研究旨在探讨H 2 S 是否过硫化转录因子FOXO1 以抑制VSMC 增殖。

方法

内皮素-1(ET-1)诱导VSMC A7r5细胞增殖后,Western blotting检测FOXO1磷酸化和增殖细胞核抗原(PCNA)表达,FOXO1核排斥程度和细胞核内PCNA荧光信号分别为通过免疫荧光检测,并通过生物素开关测定法测量 FOXO1 的过硫化。

结果

结果表明,ET-1刺激增加了细胞增殖、FOXO1磷酸化和FOXO1对细胞质的核排斥。然而,用 H 2 S 供体 NaHS 预处理成功地消除了 ET-1 诱导的 VSMC 增殖、FOXO1 磷酸化和 FOXO1 核排斥到细胞质的增加。从机制上讲,H 2 S 过硫化 A7r5 和 293T 细胞中的 FOXO1 蛋白,而硫醇还原剂 DTT 逆转了这种作用。此外,FOXO1 的 C457S 突变消除了 H 2 S 诱导的 FOXO1 在细胞中的过硫化作用以及随后对 FOXO1 Ser256 磷酸化、FOXO1 核排斥到细胞质和细胞增殖的抑制作用。

结论

因此,我们的研究结果表明,H 2 S 可能通过在 Cys457 处过硫化 FOXO1 并随后在 Ser256 处阻止 FOXO1 磷酸化来抑制 VSMC 增殖。

更新日期:2020-07-01
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