Department of Neurosurgery, Yidu Central Hospital of Weifang, Qingzhou 262500, Shandong, China.
Department of Neurosurgery, Dezhou People's Hospital, Dezhou 253014, Shandong, China.
CNS Neurol Disord Drug Targets. 2019;18(4):334-341. doi: 10.2174/1871527318666190212111650.
BACKGROUND/OBJECTIVE: Grape seed proanthocyanidins (GSPs) are a group of polyphenolic bioflavonoids, which possess a variety of biological functions and pharmacological properties. We studied the neuroprotective effects of GSP against oxygen-glucose deprivation/reoxygenation (OGD/R) injury and the potential mechanisms in mouse neuroblastoma N2a cells.
OGD/R was conducted in N2a cells. Cell viability was evaluated by CCK-8 and LDH release assay. Apoptosis was assessed by TUNEL staining and flow cytometry. Protein levels of cleaved caspase-3, Bax and Bcl-2 were detected by Western blotting. CHOP, GRP78 and caspase-12 mRNA levels were assessed by real-time PCR. JC-1 dying was used to detect mitochondrial membrane potential. ROS levels, activities of endogenous antioxidant enzymes and ATP production were examined to evaluate mitochondrial function.
GSP increased cell viability after OGD/R injury in a dose-dependent manner. Furthermore, GSP inhibited cell apoptosis, reduced the mRNA levels of CHOP, GRP78 and caspase-12 (ER stressassociated genes), restored mitochondrial membrane potential and ATP generation, improved activities of endogenous anti-oxidant ability (T-AOC, GXH-Px, and SOD), and decreased ROS level.
Our findings suggest that GSP can protect N2a cells from OGD/R insult. The mechanism of anti-apoptotic effects of GSP may involve attenuating ER stress and mitochondrial dysfunction.
背景/目的：葡萄籽油原花青素（GSP）是一组多酚类生物类黄酮，具有多种生物学功能和药理学特性。我们研究了 GSP 对氧葡萄糖剥夺/复氧（OGD/R）损伤的神经保护作用及其在小鼠神经母细胞瘤 N2a 细胞中的潜在机制。
在 N2a 细胞中进行 OGD/R。通过 CCK-8 和 LDH 释放试验评估细胞活力。通过 TUNEL 染色和流式细胞术评估细胞凋亡。通过 Western blot 检测裂解的 caspase-3、Bax 和 Bcl-2 蛋白水平。通过实时 PCR 评估 CHOP、GRP78 和 caspase-12 的 mRNA 水平。使用 JC-1 死亡来检测线粒体膜电位。检查 ROS 水平、内源性抗氧化酶活性和 ATP 生成，以评估线粒体功能。
GSP 以剂量依赖的方式增加 OGD/R 损伤后的细胞活力。此外，GSP 抑制细胞凋亡，降低 CHOP、GRP78 和 caspase-12（内质网应激相关基因）的 mRNA 水平，恢复线粒体膜电位和 ATP 生成，改善内源性抗氧化能力（T-AOC、GXH-Px 和 SOD）的活性，并降低 ROS 水平。
我们的研究结果表明，GSP 可以保护 N2a 细胞免受 OGD/R 损伤。GSP 抗凋亡作用的机制可能涉及减轻内质网应激和线粒体功能障碍。
