Rac1-NADPH oxidase-regulated generation of reactive oxygen species mediates glutamate-induced apoptosis in SH-SY5Y human neuroblastoma cells.
Reactive oxygen species (ROS) are known to play an important role in glutamate-induced neuronal cell death. In the present study, we examined whether NADPH oxidase serves as a source of ROS production and plays a role in glutamate-induced cell death in SH-SY5Y human neuroblastoma cells. Stimulation of the cells with glutamate (100 mM) induced apoptotic cell death and increase in the level of ROS, and these effects of glutamate were significantly suppressed by the inhibitors of the NADPH oxidase, diphenylene iodonium, apocynin, and neopterine. In addition, RT-PCR revealed that SH-SY5Y cells expressed mRNA of gp91phox, p22phox and cytosolic p47phox, p67phox and p40phox, the components of the plasma membrane NADPH oxidase. Treatment with glutamate also resulted in activation and translocation of Rac1 to the plasma membrane. Moreover, the expression of Rac1N17, a dominant negative mutant of Rac1, significantly blocked the glutamate-induced ROS generation and cell death. Collectively, these results suggest that the plasma membrane-bound NADPH oxidase complex may play an essential role in the glutamate-induced apoptotic cell death through increased production of ROS.
Nikolova
Sevdalina
S
Department of Biotechnology, Graduate School, Yeungnam University, Gyeongsan, 712-749, South Korea.
Lee
Yong Soo
YS
Lee
Yoon-Seok
YS
Kim
Jung-Ae
JA
D000098:Acetophenones / Q000494:pharmacology; D017209:Apoptosis; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D018698:Glutamic Acid / Q000494:pharmacology / Q000502:physiology*; D006801:Humans; D009154:Mutation; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D019798:Neopterin / Q000494:pharmacology; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D009861:Onium Compounds / Q000494:pharmacology; D021381:Protein Transport; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D020830:rac1 GTP-Binding Protein / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D000098:Acetophenones; D009861:Onium Compounds; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D018698:Glutamic Acid; D019798:Neopterin; C007517:diphenyleneiodonium; C056165:acetovanillone; D019255:NADPH Oxidases; D020830:rac1 GTP-Binding Protein