Microvesicles Derived from Nitric Oxide Synthase-Inhibited Endothelial Cells Promote Cell Dysfunction.

IF 1.8 4区医学 Q3 PERIPHERAL VASCULAR DISEASE Journal of Vascular Research Pub Date : 2024-12-10 DOI:10.1159/000542280

Vinicius P Garcia, Kelly A Stockelman, Ma'ayan V Levy, Hannah K Fandl, Anabel Goulding, Jamie G Hijmans, Samuel T Ruzzene, Auburn R Berry, Jared J Greiner, Christopher A DeSouza

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Abstract

Introduction: The aims of this study were to determine (1) whether endothelial nitric oxide synthase (eNOS) inhibition stimulates endothelial microvesicles (EMVs) release and (2) the effect of EMVs derived from eNOS-inhibited cells on endothelial cell eNOS, inflammation, apoptosis, and tissue-type plasminogen activator (t-PA).

Methods: Human umbilical vein endothelial cells (HUVECs) were treated with the eNOS inhibitor (NG-nitro-l-arginine methyl ester [L-NAME], 300 µM) for 24 h. EMVs from untreated and L-NAME-treated cells were isolated, quantified, and exposed to HUVECs for 24 h.

Results: eNOS-inhibited cells released significantly higher EMVs than untreated cells (81 ± 13 vs. 41 ± 15 EMV/μL; p = 0.005). Expression of total eNOS (97.1 ± 16.4 vs. 157.5 ± 31.2 arbitrary units [AUs]; p = 0.01), p-eNOS (4.9 ± 1.2 vs. 9.1 ± 12.6 AUs; p = 0.02), and NO production (5.0 ± 0.8 vs. 7.0 ± 1.3 µmol/L; p = 0.04) were significantly lower in cells treated with EMVs from L-NAME-treated cells. L-NAME-derived EMVs induced significantly higher IL-6 (38.3 ± 10.3 vs. 21.0 ± 3.8 pg/mL; p = 0.01) and IL-8 (38.9 ± 7.0 vs. 27.2 ± 6.2 pg/mL; p = 0.04) production concurrent with higher expression of p-NF-κB p65 (Ser536) (9.7 ± 1.6 vs. 6.1 ± 1.2 AUs; p = 0.01). Expression of activated caspase-3 was higher (9.5 ± 1.1 vs. 6.4 ± 0.4 AUs) and t-PA lower (24.2 ± 4.3 vs. 36.2 ± 8.4 AUs; p = 0.04) in cells treated with L-NAME-derived EMVs.

Conclusion: eNOS inhibition induces an increase in EMV release and an EMV phenotype with adverse cellular effects.

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一氧化氮合酶抑制内皮细胞产生的微泡促进细胞功能障碍。

本研究的目的是确定(1)内皮型一氧化氮合酶（eNOS）抑制是否刺激内皮微囊泡（emv）释放；(2)内皮型一氧化氮合酶抑制细胞产生的emv对内皮细胞eNOS、炎症、凋亡和组织型纤溶酶原激活物（t-PA）的影响。方法：用eNOS抑制剂（ng -硝基-l-精氨酸甲酯[L-NAME]， 300µM）处理人脐静脉内皮细胞（HUVECs） 24 h，分离并定量处理未处理和L-NAME处理的细胞，并将其暴露于HUVECs 24 h。结果：eNOS抑制细胞释放的EMV明显高于未处理细胞(81±13 vs 41±15 EMV/μL；P = 0.005)。总eNOS表达量（97.1±16.4 vs 157.5±31.2任意单位[au]）；p = 0.01), p- enos(4.9±1.2 vs. 9.1±12.6 au；p = 0.02)， NO产量(5.0±0.8 vs. 7.0±1.3µmol/L；p = 0.04)，用l - name处理的细胞的emv处理的细胞显著降低。l - name衍生的emv诱导IL-6显著升高（38.3±10.3 vs. 21.0±3.8 pg/mL）；p = 0.01)和IL-8(38.9±7.0∶27.2±6.2 pg/mL)；p = 0.04)产生，同时p- nf -κB p65 （Ser536）表达升高(9.7±1.6∶6.1±1.2；P = 0.01)。活化caspase-3表达较高（9.5±1.1 vs. 6.4±0.4 au）， t-PA表达较低（24.2±4.3 vs. 36.2±8.4 au）；p = 0.04)。结论：eNOS抑制诱导EMV释放增加，EMV表型具有不良细胞效应。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Vascular Research 医学-生理学

CiteScore

3.40

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： The ''Journal of Vascular Research'' publishes original articles and reviews of scientific excellence in vascular and microvascular biology, physiology and pathophysiology. The scope of the journal covers a broad spectrum of vascular and lymphatic research, including vascular structure, vascular function, haemodynamics, mechanics, cell signalling, intercellular communication, growth and differentiation. JVR''s ''Vascular Update'' series regularly presents state-of-the-art reviews on hot topics in vascular biology. Manuscript processing times are, consistent with stringent review, kept as short as possible due to electronic submission. All articles are published online first, ensuring rapid publication. The ''Journal of Vascular Research'' is the official journal of the European Society for Microcirculation. A biennial prize is awarded to the authors of the best paper published in the journal over the previous two years, thus encouraging young scientists working in the exciting field of vascular biology to publish their findings.