Department of Pharmacology, School of Medicine, Jinan University, Guangzhou 510632, China
School of International Education, Anhui Medical University, Hefei 230000, China
School of Nursing, Guangdong Pharmaceutical University, Guangzhou 510632, China
The First Affiliated Hospital of Jinan University, Guangzhou 510632
Department of Pathogen Biology and Medical Immunology, School of Basic Medicine, Ningxia Medical University, Yinchuan 750021, China
Department of Anatomy, Li Ka-shing Faculty of Medicine of The university of Hong Kong, Hong Kong 999077, China
Institute of Brain Sciences, Jinan University, Guangzhou 510632, China
: Received: 14 August 2016 / Approved: 15 August 2016 / Online: 15 August 2016 (10:42:05 CEST)
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Cai, L.; Wang, L.; Pan, J.; Mi, X.; Zhang, Z.; Geng, H.; Wang, J.; Hu, S.; Zhang, W.; Gao, Q.; Wu, W.; Luo, H. Neuroprotective Effects of Methyl 3,4-Dihydroxybenzoate Against TBHP-Induced Oxidative Damage in SH-SY5Y Cells. Preprints2016, 2016080145 (doi: 10.20944/preprints201608.0145.v1).
Cai, L.; Wang, L.; Pan, J.; Mi, X.; Zhang, Z.; Geng, H.; Wang, J.; Hu, S.; Zhang, W.; Gao, Q.; Wu, W.; Luo, H. Neuroprotective Effects of Methyl 3,4-Dihydroxybenzoate Against TBHP-Induced Oxidative Damage in SH-SY5Y Cells. Preprints 2016, 2016080145 (doi: 10.20944/preprints201608.0145.v1).
This study investigated the neuroprotective effects of methyl 3,4-dihydroxybenzoate (MDHB) against t-butylhydroperoxide(TBHP) induced oxidative damage in SH-SY5Y (human neuroblastoma cells) and the underlying mechanisms. SH-SY5Y were cultured in DMEM+10% FBS for 24 hours and pretreated with different concentrations of MDHB or N-acetyl-L-cysteine (NAC) for 4 hours prior to the addition of 40 μM TBHP for 24 hours. Cell viability was analyzed using the methyl thiazolyl tetrazolium (MTT) and lactate dehydrogenase (LDH) assays. An annexin V-FITC assay was used to detect cell apoptosis rate. 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