Cytoprotection by nipradilol, an anti-glaucomatous agent, via down-regulation of apoptosis related gene expression and activation of NF-kappaB.
It has been reported that nipradilol, a nonselective beta- and selective alpha1-receptor antagonist, has cytoprotective effects. We attempted to clarify the effects of nipradilol on the expression of apoptosis associated genes and the activity of nuclear factor-kappaB, a transcription factor, in PC12 cells during serum deprivation induced apoptosis. PC12 cells were cultured in serum free RPMI1640 medium with or without 0.01, 0.1, 1, or 10 microM of nipradilol, or in serum-added medium as a control. The gene expressions of Bax, Bcl-2, Fas, FasL, Caspase-1, 2, 3, and 9, p53, and Smac/DIABLO were examined using a quantitative real time polymerase chain reaction method, while nuclear factor-kappaB activity was examined using an electrophoresis mobility shift assay with a nuclear factor-kappaB consensus sequenced DNA probe. The effects of denitronipradilol were also examined to clarify the effect of nitric oxide donative action. Nipradilol down-regulated Bax gene expression 12 hr after serum deprivation, and that of the capase-9 and Smac/DIABLO genes at 24 hr, compared to the serum-free sample, while it also increased cell viability and decreased DNA ladder formation at 48 hr. However, the expressions of other examined genes were not affected by the agent. In addition, nuclear factor-kappaB activity was increased 2 hr after the addition of 0.1 or 1 microM of nipradilol. In contrast, denitronipradilol did not show any effects toward PC12 cells. Our results suggest that nipradilol may have an effect on apoptosis associated gene expression and nuclear factor-kappaB activity during the prevention of apoptosis via nitric oxide donative action.
Ando
Akira
A
Department of Ophthalmology, Kansai Medical University, 10-15 Fumizono, Moriguchi, Osaka 570-8507, Japan. andoak@takii.kmu.ac.jp
Yamazaki
Yukari
Y
Kaneko
Shiho
S
Miyake
Maki
M
Nambu
Rie
R
Taomoto
Makoto
M
Unezaki
Sawako
S
Okuda-Ashitaka
Emiko
E
Okumura
Tadayoshi
T
Ito
Seiji
S
Matsumura
Miyo
M
D000317:Adrenergic alpha-Antagonists / Q000494:pharmacology*; D000319:Adrenergic beta-Antagonists / Q000494:pharmacology*; D000818:Animals; D000959:Antihypertensive Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D051017:Apoptosis Regulatory Proteins; D002352:Carrier Proteins / Q000235:genetics; D053453:Caspase 9; D020169:Caspases / Q000235:genetics; D015536:Down-Regulation / Q000235:genetics; D005786:Gene Expression Regulation / Q000502:physiology*; D005901:Glaucoma / Q000517:prevention & control; D024101:Mitochondrial Proteins / Q000235:genetics; D016328:NF-kappa B / Q000378:metabolism*; D016716:PC12 Cells; D016133:Polymerase Chain Reaction / Q000379:methods; D011412:Propanolamines / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics; D051381:Rats; D014178:Translocation, Genetic / Q000235:genetics; D051028:bcl-2-Associated X Protein
D000317:Adrenergic alpha-Antagonists; D000319:Adrenergic beta-Antagonists; D000959:Antihypertensive Agents; D051017:Apoptosis Regulatory Proteins; C494828:Bax protein, rat; D002352:Carrier Proteins; C489013:DIABLO protein, rat; D024101:Mitochondrial Proteins; D016328:NF-kappa B; D011412:Propanolamines; D019253:Proto-Oncogene Proteins c-bcl-2; D051028:bcl-2-Associated X Protein; C506435:Casp9 protein, rat; D053453:Caspase 9; D020169:Caspases; C036089:nipradilol