Neuroprotective Effects of Methyl 3,4-Dihydroxybenzoate against TBHP-Induced Oxidative Damage in SH-SY5Y Cells

• Main Authors: Liang Cai, Li-Fang Wang, Jun-Ping Pan, Xiang-Nan Mi, Zheng Zhang, Hai-Ju Geng, Jia-Hui Wang, Song-Hui Hu, Wei Zhang, Qin Gao, Wu-Tian Wu, Huan-Min Luo
• Format: Article
• Language: English
• Published: MDPI AG 2016-08-01
• Series: Molecules
• Subjects: methyl 3,4-dihydroxybenzoate; oxidative stress; apoptosis; neuroprotection; nuclear factor erythroid 2-related factor 2
• Online Access: http://www.mdpi.com/1420-3049/21/8/1071

This study investigated the neuroprotective effects of methyl 3,4-dihydroxybenzoate (MDHB) against t-butyl hydroperoxide (TBHP) induced oxidative damage in SH-SY5Y (human neuroblastoma cells) and the underlying mechanisms. SH-SY5Y were cultured in DMEM + 10% FBS for 24 h and pretreated with different concentrations of MDHB or N-acetyl-l-cysteine (NAC) for 4 h prior to the addition of 40 μM TBHP for 24 h. Cell viability was analyzed using the methylthiazolyltetrazolium (MTT) and lactate dehydrogenase (LDH) assays. An annexin V-FITC assay was used to detect cell apoptosis rates. The 2′,7′-dichlorofluorescin diacetate (DCFH-DA) assay was used to determine intracellular ROS levels. The activities of antioxidative enzymes (GSH-Px and SOD) were measured using commercially available kits. The oxidative DNA damage marker 8-OHdG was detected using ELISA. Western blotting was used to determine the expression of Bcl-2, Bax, caspase 3, p-Akt and Akt proteins in treated SH-SY5Y cells. Our results showed that MDHB is an effective neuroprotective compound that can mitigate oxidative stress and inhibit apoptosis in SH-SY5Y cells.