Student Research Committee, Birjand University of Medical Sciences, Birjand, Iran.
Department of Medical Immunology, Faculty of Medicine, Birjand University of Medical Sciences, Birjand, Iran.
Mediators Inflamm. 2022 Aug 31;2022:5171525. doi: 10.1155/2022/5171525. eCollection 2022.
Inflammation is the body's biological reaction to endogenous and exogenous stimuli. Recent studies have demonstrated several anti-inflammatory properties of species. In this paper, we decided to study the anti-inflammatory effect of ethanolic extract of oleo-gum-resin (asafoetida) against TNF--stimulated human umbilical vein endothelial cells (HUVECs). HUVECs were cultured in a flat-bottom plate and then treated with ethanolic extract of asafoetida (EEA, 0-500 g/ml) and TNF- (0-100 ng/ml) for 24 h. We used the MTT test to assess cell survival. In addition, the LC-MS analysis was performed to determine the active substances. HUVECs were pretreated with EEA and then induced by TNF-. Intracellular reactive oxygen species (ROS) and adhesion of peripheral blood mononuclear cells (PBMCs) to HUVECs were evaluated with DCFH-DA and CFSE fluorescent probes, respectively. Gene expression of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin and surface expression of ICAM-1 protein were measured using real-time PCR and flow cytometry methods, respectively. While TNF- significantly increased intracellular ROS formation and PBMC adhesion to TNF--induced HUVECs, the pretreatment of HUVECs with EEA (125 and 250 g/ml) significantly reduced the parameters. In addition, EEA pretreatment decreased TNF--induced mRNA expression of VCAM-1 and surface protein expression of ICAM-1 in the target cells. Taken together, the results indicated that EEA prevented ROS generation, triggered by TNF-, and inhibited the expression of VCAM-1 and ICAM-1, leading to reduced PBMC adhesion. These findings suggest that EEA can probably have anti-inflammatory properties.
炎症是机体对内外源性刺激的生物学反应。最近的研究表明，[物种名称]具有多种抗炎特性。在本文中，我们决定研究[植物名称]乙醇提取物（阿魏胶）对 TNF-刺激的人脐静脉内皮细胞（HUVEC）的抗炎作用。将 HUVEC 培养在平底培养板中，然后用[植物名称]乙醇提取物（EEA，0-500μg/ml）和 TNF-（0-100ng/ml）处理 24 小时。我们使用 MTT 试验来评估细胞存活率。此外，还进行了 LC-MS 分析以确定活性物质。用 EEA 预处理 HUVEC，然后用 TNF-诱导。用 DCFH-DA 和 CFSE 荧光探针分别评估细胞内活性氧（ROS）和外周血单核细胞（PBMC）与 HUVEC 的黏附。使用实时 PCR 和流式细胞术方法分别测量细胞间黏附分子-1（ICAM-1）、血管细胞黏附分子 1（VCAM-1）和 E-选择素的基因表达以及 ICAM-1 蛋白的表面表达。虽然 TNF-显著增加了细胞内 ROS 的形成和 PBMC 与 TNF-诱导的 HUVEC 的黏附，但 EEA（125 和 250μg/ml）预处理显著降低了这些参数。此外，EEA 预处理降低了 TNF-诱导的靶细胞中 VCAM-1 的 mRNA 表达和 ICAM-1 的表面蛋白表达。综上所述，这些结果表明，EEA 可预防 TNF-引发的 ROS 生成，并抑制 VCAM-1 和 ICAM-1 的表达，从而减少 PBMC 的黏附。这些发现表明，EEA 可能具有抗炎特性。
