P物质通过体外调节内皮型一氧化氮合酶（eNOS）的表达改善肿瘤坏死因子-α诱导的内皮细胞功能障碍。

Substance P ameliorates tumor necrosis factor-alpha-induced endothelial cell dysfunction by regulating eNOS expression in vitro.

作者信息

Piao Jiyuan, Hong Hyun Sook, Son Youngsook

机构信息

Department of Genetic Engineering, College of Life Science and Graduate School of Biotechnology, Kyung Hee University, Yong In, Korea.

East-West Medical Research Institute/Department of Biomedical Science and Technology, Graduate School, Kyung Hee University, Seoul, Korea.

出版信息

Microcirculation. 2018 Apr;25(3):e12443. doi: 10.1111/micc.12443.

DOI:10.1111/micc.12443

PMID:29412499

Abstract

OBJECTIVE

The aim of this study was to explore the beneficial effects of SP on NO production and inflammation-induced vascular endothelium cell death.

METHODS

To mimic the inflammatory environment, TNF-α was treated with HUVECs, and SP was added prior to TNF-α to determine its protective effect. WST-1 assay was performed to detect cell viability. NO level in conditioned medium was measured by Griess Reagent System. The protein level of cleaved caspase-3, eNOS, and phosphorylated Akt was detected by Western blot analysis.

RESULTS

TNF-α declined endothelial cell viability by downregulating Akt and NO production. TNF-α-induced cell death was reliably restored by NO, confirming the requirement of NO for cell survival. By contrast, pretreatment of SP attenuated TNF-α-induced cellular apoptosis, accompanied by an increase in the phosphorylation of Akt, eNOS expression, and NO production. Blockage of NK-1R, phosphorylated Akt or eNOS by CP-96345, A6730, or L-NAME entirely eliminated the effect of SP.

CONCLUSIONS

SP can protect the vascular endothelium against inflammation-induced damage through modulation of the Akt/eNOS/NO signaling pathway.

摘要

目的

本研究旨在探讨速激肽（SP）对一氧化氮（NO）生成及炎症诱导的血管内皮细胞死亡的有益作用。

方法

为模拟炎症环境，用肿瘤坏死因子-α（TNF-α）处理人脐静脉内皮细胞（HUVECs），并在加入TNF-α之前添加SP以确定其保护作用。采用WST-1法检测细胞活力。用格里斯试剂系统测定条件培养基中的NO水平。通过蛋白质印迹分析检测裂解的半胱天冬酶-3、内皮型一氧化氮合酶（eNOS）和磷酸化Akt的蛋白质水平。

结果

TNF-α通过下调Akt和NO生成降低内皮细胞活力。NO可可靠地恢复TNF-α诱导的细胞死亡，证实NO对细胞存活的必要性。相比之下，SP预处理可减轻TNF-α诱导的细胞凋亡，同时伴有Akt磷酸化增加、eNOS表达增加和NO生成增加。CP-96345、A6730或L-NAME阻断神经激肽-1受体（NK-1R）、磷酸化Akt或eNOS可完全消除SP的作用。