鹿茸多肽（VAP）通过体外 NF-κB 信号通路保护脑缺血损伤。

Velvet antler polypeptide (VAP) protects against cerebral ischemic injury through NF-κB signaling pathway in vitro.

机构信息

Physical Examination Center, Wuhan Third Hospital, Tongren Hospital of Wuhan University, Wuhan 430074, Hubei Province, China.

出版信息

J Stroke Cerebrovasc Dis. 2024 May;33(5):107666. doi: 10.1016/j.jstrokecerebrovasdis.2024.107666. Epub 2024 Feb 27.

DOI:10.1016/j.jstrokecerebrovasdis.2024.107666

PMID:38423152

Abstract

OBJECTIVE

Velvet antler polypeptide (VAP) has been shown to play important roles in the immune and nervous systems. The purpose of this study was to investigate the protective effects of VAP on cerebral ischemic injury with the involvement of NF-κB signaling pathway in vitro.

MATERIALS AND METHODS

PC-12 cells stimulated by oxygen-glucose deprivation/reperfusion (OGD/R) was used to mimic cerebral ischemic injury in vitro. The levels of ROS, SOD, and intracellular concentrations of Ca2+ were measured by the relevant kits. Meanwhile, the expressions of inflammatory cytokines (IL-6, IL-1β, and TNF-α) were determined by ELISA kit assay. In addition, MTT, EdU, and flow cytometry assays were used to measure the cell proliferation and apoptosis. Besides which, the related proteins of NF-κB signaling pathway were measured by western blotting assay.

RESULTS

VAP alleviated cerebral ischemic injury by reducing OGD/R-induced oxidative stress, inflammation, and apoptosis in PC-12 cells in a time dependent manner. Mechanistically, VAP inhibited the levels of p-p65 and p-IkB-α in a time dependent manner, which was induced by OGD/R operation. Moreover, NF-κB agonist diprovocim overturned the suppression effects of VAP on OGD/R-induced oxidative stress, inflammation, and apoptosis in PC-12 cells.

CONCLUSIONS

The results demonstrate that VAP may alleviate cerebral ischemic injury by suppressing the activation of NF-κB signaling pathway.

摘要

目的

鹿茸多肽（VAP）已被证明在免疫系统和神经系统中发挥重要作用。本研究旨在探讨 VAP 通过 NF-κB 信号通路对体外脑缺血损伤的保护作用。

材料和方法

采用氧葡萄糖剥夺/再灌注（OGD/R）刺激的 PC-12 细胞体外模拟脑缺血损伤。通过相关试剂盒测量 ROS、SOD 和细胞内 Ca2+浓度。同时，通过 ELISA 试剂盒测定炎症细胞因子（IL-6、IL-1β和 TNF-α）的表达。此外，通过 MTT、EdU 和流式细胞术测定细胞增殖和凋亡。此外，通过 Western blot 测定 NF-κB 信号通路的相关蛋白。

结果

VAP 通过减轻 OGD/R 诱导的 PC-12 细胞氧化应激、炎症和凋亡，从而减轻脑缺血损伤，且呈时间依赖性。机制上，VAP 抑制了 OGD/R 诱导的 p-p65 和 p-IkB-α的水平，呈时间依赖性。此外，NF-κB 激动剂二丙伏昔可逆转 VAP 对 OGD/R 诱导的 PC-12 细胞氧化应激、炎症和凋亡的抑制作用。