Department of Orthopedics, Lanzhou University Second Hospital, Lanzhou, Gansu, 730000, China; Orthopaedics Key Laboratory of Gansu Province, Lanzhou, Gansu, 730000, China.
General Practice Medicine, Yanan University Affiliated Hospital, Yanan, Shaanxi 716000, China.
Neurosci Lett. 2021 Apr 17;750:135793. doi: 10.1016/j.neulet.2021.135793. Epub 2021 Mar 2.
Grape seed proanthocyanidins (GSP) are natural flavonoids with strong antioxidant and anti-apoptotic effects. Oxidative stress and neuronal apoptosis are major contributors to spinal cord injury (SCI). In this study, we assessed the potential protective effects of GSP on hydrogen peroxide (HO)-damaged pheochromocytoma-12 (PC12) cells in an in vitro model of SCI as well as the putative mechanism of action. We established a model using PC12 cells with oxidative damage induced by HO. Cells were treated with various concentrations of GSP (control group, 200 μmol/L HO group, 5 μM GSP + HO group, 10 μM GSP + HO group, and 25 μM GSP + HO group). The CCK-8 assay was used to determine cell activity. Dichloro-dihydro-fluorescein diacetate was used to detect intracellular reactive oxygen species (ROS), and flow cytometry was used to determine apoptosis rate. Western blot analysis was used to detect the expression of caspase-3, Bax, Bcl-2, and PI3K/AKT proteins. The results showed that GSP reduced HO-induced intracellular ROS and inhibited apoptosis. Furthermore, GSP inhibited the expression of caspase-3 and Bax, while promoting the expression of Bcl-2. In addition, GSP promoted the phosphorylation of PI3K and AKT. Moreover, a PI3K inhibitor (LY294002) weakened the protective effects of GSP on HO-induced PC12 cells. In conclusion, GSP pretreatment can protect PC12 cells from oxidative damage induced by HO via the PI3K/AKT signaling pathway.
葡萄籽原花青素（GSP）是一种具有强抗氧化和抗凋亡作用的天然类黄酮。氧化应激和神经元凋亡是导致脊髓损伤（SCI）的主要因素。在本研究中，我们评估了 GSP 在体外 SCI 模型中对过氧化氢（HO）损伤的嗜铬细胞瘤-12（PC12）细胞的潜在保护作用及其可能的作用机制。我们建立了一个使用 HO 诱导的氧化损伤的 PC12 细胞模型。用不同浓度的 GSP（对照组、200μmol/L HO 组、5μM GSP+HO 组、10μM GSP+HO 组和 25μM GSP+HO 组）处理细胞。使用 CCK-8 测定法测定细胞活性。使用二氯二氢荧光素二乙酸酯检测细胞内活性氧（ROS），并使用流式细胞术测定细胞凋亡率。使用 Western blot 分析检测 caspase-3、Bax、Bcl-2 和 PI3K/AKT 蛋白的表达。结果表明，GSP 降低了 HO 诱导的细胞内 ROS 并抑制了细胞凋亡。此外，GSP 抑制了 caspase-3 和 Bax 的表达，同时促进了 Bcl-2 的表达。此外，GSP 促进了 PI3K 和 AKT 的磷酸化。此外，PI3K 抑制剂（LY294002）削弱了 GSP 对 HO 诱导的 PC12 细胞的保护作用。总之，GSP 预处理可以通过 PI3K/AKT 信号通路保护 PC12 细胞免受 HO 诱导的氧化损伤。
