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 . PC-12 cells stimulated by oxygen-glucose deprivation/reperfusion (OGD/R) was used to mimic cerebral ischemic injury . 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. 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. The results demonstrate that VAP may alleviate cerebral ischemic injury by suppressing the activation of NF-κB signaling pathway.

中文翻译：

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鹿茸多肽（VAP）通过NF-κB信号通路发挥体外保护脑缺血损伤作用

鹿茸多肽（VAP）已被证明在免疫和神经系统中发挥重要作用。本研究旨在探讨VAP通过NF-κB信号通路对脑缺血损伤的保护作用。通过氧-葡萄糖剥夺/再灌注（OGD/R）刺激的PC-12细胞被用来模拟脑缺血损伤。采用相关试剂盒测定ROS、SOD水平和细胞内Ca2+浓度。同时，采用ELISA试剂盒测定炎症细胞因子（IL-6、IL-1β、TNF-α）的表达。此外，使用MTT、EdU和流式细胞术测定细胞增殖和凋亡。此外，通过Western blotting检测NF-κB信号通路的相关蛋白。VAP 通过以时间依赖性方式减少 OGD/R 诱导的 PC-12 细胞氧化应激、炎症和细胞凋亡来减轻脑缺血损伤 从机制上讲，VAP 以时间依赖性方式抑制 p-p65 和 p-IkB-α 的水平，这是由 OGD/R 操作引起的。此外，NF-κB 激动剂 diprovocim 推翻了 VAP 对 OGD/R 诱导的 PC-12 细胞氧化应激、炎症和细胞凋亡的抑制作用。结果表明VAP可能通过抑制NF-κB信号通路的激活来减轻脑缺血损伤。