Guanosine protects human neuroblastoma SH-SY5Y cells against mitochondrial oxidative stress by inducing heme oxigenase-1 via PI3K/Akt/GSK-3¦Â pathway

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• 作者：Tharine Dal-Cim ; Simone Molz ; Javier Egea ; Esther Parada ; Alej ; ro Romero ; Josiane Budni ; Maria D. Mart¨ªn de Saavedra ; Laura del Barrio ; Carla I. Tasca ; Manuela G. L¨®pez
• 关键词：BK ; large (big) conductance Ca2+-activated K+ channels ; DPCPX ; 1 ; 3-dipropyl-8-cyclopentylxanthine ; GUO ; guanosine ; GSK-3¦Â ; glycogen synthase kinase 3¦Â ; H2DCFDA ; 2&prime ; 7&prime ; -dichlorofluorescein diacetate ; HO-1 ; heme oxygenase-1 ; LY294002 ; 2-(4-morp
• 刊名：Neurochemistry International
• 出版年：2012
• 出版时间：August, 2012
• 年：2012
• 卷：61
• 期：3
• 页码：397-404
• 全文大小：759 K

文摘

Mitochondrial perturbation and oxidative stress are key factors in neuronal vulnerability in several neurodegenerative diseases or during brain ischemia. Here we have investigated the protective mechanism of action of guanosine, the guanine nucleoside, in a human neuroblastoma cell line, SH-SY5Y, subjected to mitochondrial oxidative stress. Blockade of mitochondrial complexes I and V with rotenone plus oligomycin (Rot/oligo) caused a significant decrease in cell viability and an increase in ROS production. Guanosine that the protective effect of guanosine incubated concomitantly with Rot/oligo abolished Rot/oligo-induced cell death and ROS production in a concentration dependent manner; maximum protection was achieved at the concentration of 1 mM. The cytoprotective effect afforded by guanosine was abolished by adenosine A₁ or A_2A receptor antagonists (DPCPX or ZM241385, respectively), or by a large (big) conductance Ca²⁺-activated K⁺ channel (BK) blocker (charybdotoxin). Evaluation of signaling pathways showed that the protective effect of guanosine was not abolished by a MEK inhibitor (PD98059), by a p38^MAPK inhibitor (SB203580), or by a PKC inhibitor (cheleritrine). However, when blocking the PI3K/Akt pathway with LY294002, the neuroprotective effect of guanosine was abolished. Guanosine increased Akt and p-Ser-9-GSK-3¦Â phosphorylation confirming this pathway plays a key role in guanosine¡¯s neuroprotective effect. Guanosine induced the antioxidant enzyme heme oxygenase-1 (HO-1) expression. The protective effects of guanosine were prevented by heme oxygenase-1 inhibitor, SnPP. Moreover, bilirubin, an antioxidant and physiologic product of HO-1, is protective against mitochondrial oxidative stress. In conclusion, our results show that guanosine can afford protection against mitochondrial oxidative stress by a signaling pathway that implicates PI3K/Akt/GSK-3¦Â proteins and induction of the antioxidant enzyme HO-1.