Wu Shuang, Yang Tiantian, Cen Kai, Zou Yihuai, Shi Xiaowei, Zhou Dongrui, Gao Yonghong, Chai Limin, Zhao Yizhou, Sun Yikun, Zhu Lingqun
Key Laboratory of Chinese Internal Medicine of Educational Ministry and Beijing, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
Department of Stomatology, Dongzhimen Hospital, Beijing University of Chinese Medicine, Beijing, China.
Evid Based Complement Alternat Med. 2020 Jul 31;2020:1403572. doi: 10.1155/2020/1403572. eCollection 2020.
This study investigated whether saponins (PNS) extracted from (Bruk.) F. H. Chen played a neuroprotective role by affecting the EGFR/PI3K/AKT pathway in oxygen-glucose deprived (OGD) SH-SY5Y cells.
Different groups of OGD SH-SY5Y cells were treated with varying doses of PNS, PNS + AG1478 (a specific inhibitor of EGFR), or AG1478 for 16 hours. CCK8, Annexin V-FITC/PI apoptosis analysis, and LDH release analysis were used to determine cell viability, apoptosis rate, and amounts of LDH. Quantitative real-time PCR (q-RT-PCR) and western blotting were used to measure mRNA and proteins levels of p-EGFR/EGFR, p-PI3K/PI3K, and p-AKT/AKT in SH-SY5Y cells subjected to OGD.
PNS significantly enhanced cell viability, reduced apoptosis, and weakened cytotoxicity by inhibiting the release of LDH. The mRNA expression profiles of EGFR, PI3K, and AKT showed no difference between model and other groups. Additionally, ratios of p-EGFR, p-PI3K, and p-AKT to EGFR, PI3K, and AKT proteins expression, respectively, all increased significantly.
These findings indicate that PNS enhanced neuroprotective effects by activating the EGFR/PI3K/AKT pathway and elevating phosphorylation levels in OGD SH-SY5Y cells.
本研究探讨从三七（Bruk.）F. H. Chen中提取的皂苷（PNS）是否通过影响氧糖剥夺（OGD）的SH-SY5Y细胞中的表皮生长因子受体（EGFR）/磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（AKT）信号通路发挥神经保护作用。
将不同组的OGD SH-SY5Y细胞用不同剂量的PNS、PNS + AG1478（EGFR的特异性抑制剂）或AG1478处理16小时。采用细胞计数试剂盒-8（CCK8）、膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）凋亡分析和乳酸脱氢酶（LDH）释放分析来测定细胞活力、凋亡率和LDH含量。采用定量实时聚合酶链反应（q-RT-PCR）和蛋白质印迹法来检测OGD处理的SH-SY5Y细胞中磷酸化EGFR/EGFR、磷酸化PI3K/PI3K和磷酸化AKT/AKT的mRNA和蛋白水平。
PNS通过抑制LDH释放显著提高细胞活力、减少凋亡并减弱细胞毒性。模型组与其他组之间EGFR、PI3K和AKT的mRNA表达谱无差异。此外，磷酸化EGFR、磷酸化PI3K和磷酸化AKT与EGFR、PI3K和AKT蛋白表达的比值均显著增加。
这些结果表明，PNS通过激活OGD处理的SH-SY5Y细胞中的EGFR/PI3K/AKT信号通路并提高磷酸化水平来增强神经保护作用。
