Ramalingam Mahesh, Kim Sung-Jin
Department of Pharmacology and Toxicology, Metabolic Diseases Research Laboratory, School of Dentistry, Kyung Hee University , Seoul , Republic of Korea.
J Recept Signal Transduct Res. 2014 Jun;34(3):212-20. doi: 10.3109/10799893.2013.876043. Epub 2014 Jan 24.
Exogenous hydrogen peroxide (H2O2) can easily penetrate into biological membranes and enhance the formation of other reactive oxygen species (ROS). In the present study, we have investigated the neuroprotective effects of insulin on H2O2-induced toxicity of retinoic acid (RA)-differentiated SH-SY5Y cells. To measure the changes in the cell viability of SH-SY5Y cells at different concentrations of H2O2 for 24 h, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT)-based assay was used and a 100 µM H2O2 was selected to establish a model of H2O2-induced oxidative stress. Further assays showed that 24 h of 100 µM H2O2-induced significant changes in the levels of lactate dehydrogenase (LDH), nitric oxide (NO), ROS, and calcium ion (Ca2+) in neuronal cells, but insulin can effectively diminish the H2O2-induced oxidative damages to these cells. Moreover, cells treated with insulin increased H2O2-induced suppression of glutathione levels and exerted an apparent suppressive effect on oxidative products. The results of insulin treatment with SH-SY5Y cells increased the Bcl-2 levels and decreased the Akt levels. The treatment of insulin had played a protective effect on H2O2-induced oxidative stress related to the Akt/Bcl-2 pathways.
外源性过氧化氢（H2O2）能够轻易穿透生物膜并促进其他活性氧（ROS）的形成。在本研究中，我们探究了胰岛素对H2O2诱导的视黄酸（RA）分化的SH-SY5Y细胞毒性的神经保护作用。为了测定不同浓度的H2O2作用24小时后SH-SY5Y细胞活力的变化，采用了基于3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）的检测方法，并选择100µM H2O2建立H2O2诱导的氧化应激模型。进一步的检测表明，100µM H2O2作用24小时会导致神经元细胞中乳酸脱氢酶（LDH）、一氧化氮（NO）、ROS和钙离子（Ca2+）水平发生显著变化，但胰岛素可有效减轻H2O2对这些细胞的氧化损伤。此外，用胰岛素处理的细胞增加了H2O2诱导的谷胱甘肽水平抑制，并对氧化产物产生明显的抑制作用。用胰岛素处理SH-SY5Y细胞的结果增加了Bcl-2水平并降低了Akt水平。胰岛素处理对与Akt/Bcl-2途径相关的H2O2诱导的氧化应激起到了保护作用。
