Wang Meng-xia, Zhao Jing-yu, Sun Dong-mei, Meng Xiang-bao, Sun Gui-bo, Sun Xiao-bo
Yao Xue Xue Bao. 2016 Jun;51(6):898-906.
The aim of this study is to investigate the protective effects of Panax notoginseng saponins (PNS) against 6-hydroxydopamine(6-OHDA)-induced apoptosis in SH-SY5Y cells and the possible underlying mechanisms. Cell viability was examined by MTT assay. The levels of lactate dehydrogenase(LDH), reactive oxygen species (ROS), malondialdehyde (MDA) and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase(GSH-Px) were measured using the respective assay kits. Apoptosis was measured by TUNEL kit, JC-1 and ROS was measured by staining with fluorescent dyes. The activation of caspase-3 was measured with the caspase-3 assay kit. The expression of nuclear protein Nrf2 and HO-1 were determined by Western blot. PNS had significant protective effects against 6-OHDA-induced apoptosis in SH-SY5Y cells in a time- and dose-dependent manner. PNS could attenuate 6-OHDA-induced suppression of SOD, GAT, GSH-Px (P < 0.01). PNS reduced the level of LDH, decreased the levels of ROS, MDA and increased cell viability and the mitochondrial membrane potential (P < 0.01). PNS also inhibited DNA fragmentation, mitochondrial response and the activation of caspase-3 (P < 0.01). Moreover, PNS pretreatment increased the expression of the nuclear Nrf2 and up-regulate HO-1. The protective effects of PNS could be inhibited by HO-1 inhibitor SnPP. In conclusion, PNS has significant protective effects against 6-OHDA- induced apoptosis in SH-SY5Y cells. The possible mechanisms of PNS are due to PNS-mediated activation of Nrf2, up-regulation of HO-1 and inhibition of oxidative stress.
本研究旨在探讨三七总皂苷（PNS）对6-羟基多巴胺（6-OHDA）诱导的SH-SY5Y细胞凋亡的保护作用及其可能的潜在机制。采用MTT法检测细胞活力。使用相应的检测试剂盒测定乳酸脱氢酶（LDH）、活性氧（ROS）、丙二醛（MDA）水平以及超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽过氧化物酶（GSH-Px）的活性。通过TUNEL试剂盒检测细胞凋亡，用JC-1检测线粒体膜电位，用荧光染料染色检测ROS。用caspase-3检测试剂盒检测caspase-3的激活情况。通过蛋白质免疫印迹法测定核蛋白Nrf2和HO-1的表达。PNS对6-OHDA诱导的SH-SY5Y细胞凋亡具有显著的保护作用，且呈时间和剂量依赖性。PNS可减轻6-OHDA诱导的SOD、GAT、GSH-Px的抑制作用（P<0.01）。PNS降低了LDH水平，降低了ROS、MDA水平，提高了细胞活力和线粒体膜电位（P<0.01）。PNS还抑制了DNA片段化、线粒体反应和caspase-3的激活（P<0.01）。此外，PNS预处理可增加核Nrf2的表达并上调HO-1。HO-1抑制剂SnPP可抑制PNS的保护作用。综上所述，PNS对6-OHDA诱导的SH-SY5Y细胞凋亡具有显著的保护作用。PNS的可能机制是由于其介导的Nrf2激活、HO-1上调以及氧化应激抑制。
