Department of Physiology and Pathophysiology, National Key Discipline of Cell Biology, Air Force Medical University, No. 169 West Changle Road, Xi'an 710032, Shaanxi Province, People's Republic of China.
School of Life Sciences, Northwest University, No.1 North Taibai Road, Xi'an 710069, Shaanxi Province, People's Republic of China.
Cytokine. 2021 Oct;146:155659. doi: 10.1016/j.cyto.2021.155659. Epub 2021 Jul 28.
The current study aims to investigate the effect of κ-opioid receptor (κ-OR) activation on sodium palmitate (SP)-induced human umbilical vein endothelial cells (HUVECs) inflammatory response and elucidate the underlying mechanisms.
A hyperlipidemic cell model was established and treated with κ-OR agonist (U50,488H), and antagonist (norbinaltorphimine, nor-BNI), or inhibitors targeting PI3K, Akt or eNOS (LY294002, MK2206-2HCl or L-NAME, respectively). Furthermore, the expression levels of NLRP3, caspase-1, p-Akt, Akt, p-eNOS, and total eNOS were evaluated. Additionally, the production of reactive oxygen species, and levels of inflammatory factors, such as TNF-α, IL-1β, IL-6, IL-1 and adhesion molecules, such as ICAM-1, VCAM-1, P-selectin, and E-selectin were determined. The adherence rates of the neutrophils and monocytes were assessed as well.
The SP-induced hyperlipidemic cell model demonstrated increased expression of NLRP3 and caspase-1 proteins (P < 0.05) and elevated ROS levels (P < 0.01), and decreased phosphorylated-Akt and phosphorylated-eNOS expression (P < 0.05). In addition, SP significantly increased TNF-α, IL-1β, IL-6, ICAM-1, VCAM-1, P-selectin, and E-selectin levels (P < 0.01), decreased IL-10 levels (P < 0.01), and increased the adhesion rates of monocytes and neutrophils (P < 0.01). The SP-induced inflammatory response in HUVECs was ameliorated by κ-OR agonist, U50,488H. However, the protective effect of U50,488H was abolished by κ-OR antagonist, nor-BNI, and inhibitors of PI3K, Akt and eNOS.
Our findings suggest that κ-OR activation inhibits SP-induced inflammation by activating the PI3K/Akt/eNOS signaling pathway.
本研究旨在探讨 κ 阿片受体（κ-OR）激活对棕榈酸钠（SP）诱导的人脐静脉内皮细胞（HUVEC）炎症反应的影响，并阐明其潜在机制。
建立高脂血症细胞模型，并给予 κ-OR 激动剂（U50,488H）、拮抗剂（norbinaltorphimine，nor-BNI）或针对 PI3K、Akt 或 eNOS 的抑制剂（LY294002、MK2206-2HCl 或 L-NAME）处理。此外，还评估了 NLRP3、caspase-1、p-Akt、Akt、p-eNOS 和总 eNOS 的表达水平。此外，还测定了活性氧（ROS）的产生以及 TNF-α、IL-1β、IL-6、IL-1 等炎症因子和 ICAM-1、VCAM-1、P-选择素、E-选择素等粘附分子的水平。还评估了中性粒细胞和单核细胞的粘附率。
SP 诱导的高脂血症细胞模型中，NLRP3 和 caspase-1 蛋白表达增加（P<0.05），ROS 水平升高（P<0.01），磷酸化-Akt 和磷酸化-eNOS 表达降低（P<0.05）。此外，SP 显著增加了 TNF-α、IL-1β、IL-6、ICAM-1、VCAM-1、P-选择素和 E-选择素水平（P<0.01），降低了 IL-10 水平（P<0.01），并增加了单核细胞和中性粒细胞的粘附率（P<0.01）。κ-OR 激动剂 U50,488H 可改善 HUVEC 中由 SP 诱导的炎症反应。然而，κ-OR 拮抗剂 nor-BNI 和 PI3K、Akt 和 eNOS 的抑制剂消除了 U50,488H 的保护作用。
我们的研究结果表明，κ-OR 激活通过激活 PI3K/Akt/eNOS 信号通路抑制 SP 诱导的炎症反应。
