Glyceraldehyde-3-Phosphate Dehydrogenase–Monoamine Oxidase B-Mediated Cell Death-Induced by Ethanol is Prevented by Rasagiline and 1-R-Aminoindan

Abstract

The inhibitors of monoamine oxidase B (MAO B) are effectively used as therapeutic drugs for neuropsychiatric and neurodegenerative diseases. However, their mechanism of action is not clear, since the neuroprotective effect of MAO B inhibitors is associated with the blockage of glyceraldehyde-3-phosphate dehydrogenase (GAPDH)-death cascade, rather than the inhibition of MAO B. Here, we provide evidence that GAPDH potentiates the ethanol-induced activity of MAO B and brain cell toxicity. The levels of nuclear GAPDH and MAO B activity are significantly increased in brain-derived cell lines upon 75 mM ethanol-induced cell death. Over-expression of GAPDH in cells enhances ethanol-induced cell death, and also increases the ethanol-induced activation of MAO B. In contrast, the MAO B inhibitors rasagiline and selegiline (0.25 nM) and the rasagiline metabolite, 1-R-aminoindan (1 μM) decreases the ethanol-induced MAO B, prevents nuclear translocation of GAPDH and reduces cell death. In addition, GAPDH interacts with transforming growth factor-beta-inducible early gene (TIEG2), a transcriptional activator for MAO B, and this interaction is increased in the nucleus by ethanol but reduced by MAO B inhibitors and 1-R-aminoindan. Furthermore, silencing TIEG2 using RNAi significantly reduces GAPDH-induced MAO B upregulation and neurotoxicity. In summary, ethanol-induced cell death, attenuated by MAO NIH Public Access

Mechanistic Role for a Novel Glucocorticoid-KLF11 (TIEG2) Protein Pathway in Stress-induced Monoamine Oxidase A Expression

“…2Aa) (35) and siRNA-mediated KLF11 knockdown (Fig. 2Ba) (34) have been confirmed by Western blot analysis. We find that MAO A mRNA levels in KLF11-transfected samples were increased significantly ϳ2-fold compared with pcDNAtransfected cells (p Ͻ 0.02, Fig.…”

“…Reverse transcription was performed using the SuperScript III first-strand synthesis system (Invitrogen). Resulting cDNA was quantified using an iCycler MyiQ real-time PCR detection system (Bio-Rad), as described previously (22,34). Specific primers for human, mouse, and rat were used and designed as follows: human MAO A, 5Ј-CGTGATCGGAGGTGGCATTTC-3Ј (sense) and 5Ј-AAAGGCGCCCCGAAAGG-3Ј (antisense); human KLF11, 5Ј-CCTGTTGCGGATAAGACCCCTCAC-3Ј (sense) and 5Ј-AAAGCCGGCAATCTGGAGTCTGGA-3Ј (antisense); mouse MAO A, 5Ј-CCGCTCCTTTTCCATGCCTCTCAA-3Ј (sense) and 5Ј-CCCTTGTACCGCCCCTTGACTGAA-3Ј (antisense); rat MAO A, 5Ј-ACGCTCAGGAATGGGAC-AAGATG-3Ј (sense) and 5Ј-CCCACACTGCCTCACATACC-ACA-3Ј (antisense); rat KLF11, 5Ј-CTCCTGCAGGGCCGTG-ATGAC-3Ј (sense) and 5Ј-GGGGAACAGGCCACCAGAC-TTG-3Ј (antisense).…”

“…Specific primers for human, mouse, and rat were used and designed as follows: human MAO A, 5Ј-CGTGATCGGAGGTGGCATTTC-3Ј (sense) and 5Ј-AAAGGCGCCCCGAAAGG-3Ј (antisense); human KLF11, 5Ј-CCTGTTGCGGATAAGACCCCTCAC-3Ј (sense) and 5Ј-AAAGCCGGCAATCTGGAGTCTGGA-3Ј (antisense); mouse MAO A, 5Ј-CCGCTCCTTTTCCATGCCTCTCAA-3Ј (sense) and 5Ј-CCCTTGTACCGCCCCTTGACTGAA-3Ј (antisense); rat MAO A, 5Ј-ACGCTCAGGAATGGGAC-AAGATG-3Ј (sense) and 5Ј-CCCACACTGCCTCACATACC-ACA-3Ј (antisense); rat KLF11, 5Ј-CTCCTGCAGGGCCGTG-ATGAC-3Ј (sense) and 5Ј-GGGGAACAGGCCACCAGAC-TTG-3Ј (antisense). The 18 S ribosomal RNA primer was used as an internal control (22,34).…”

“…Briefly, siPORT amine transfection agent and Opti-MEM I medium were mixed with each siRNA for 10 min with a final siRNA concentration of 20 nM per 10-cm dish. The siRNA⅐siPORT amine transfection agent complex was directly added to the cell culture medium (34).…”

Discordant effects of rasagiline doses in Parkinson disease

“…6 under oxidative stress, GaPDH is inactivated, causing a switch of glucose metabolism to the pentose phosphate pathway. this increases the production of naDPH, which is required for the recycling of glutathione.…”