Apoptotic cell death triggered by nitric oxide in pancreatic beta-cells.
Nitric oxide (NO) is believed to be an effector molecule that mediates interleukin (IL)-1 beta-induced destruction and dysfunction of pancreatic beta-cells. We have demonstrated that both exogenous NO and NO generated endogenously by IL-1 beta brought about apoptosis of isolated rat pancreatic islet cells as well as pancreatic beta-cell tumor-derived cell line HIT. This apoptosis was characterized by cleavage of DNA into nucleosomal fragments of 180-200 bp and morphologically by nuclear shrinkage, chromatic condensation, and apoptotic body formation. The IL-1 beta-induced internucleosomal DNA cleavage occurred in a time- and dose-dependent manner. Actinomycin D, cycloheximide, and nitric oxide synthase inhibitors inhibited the DNA cleavage, which was correlated with the amount of NO produced, indicating that NO produced by HIT cells themselves could mediate the apoptosis. Furthermore, in the presence of tumor necrosis factor (TNF)-alpha, large amounts of NO were produced by IL-1 beta and DNA cleavage occurred more noticeably, although TNF-alpha alone did not generate NO. Streptozotocin (STZ), a diabetogenic reagent containing a nitroso moiety, also released NO and induced internucleosomal DNA cleavage in HIT cells. These results suggest that NO-induced internucleosomal DNA cleavage is an important initial step in the destruction and dysfunction of pancreatic beta-cells induced by inflammatory stimulation or treatment with STZ.
Kaneto
H
H
Department of Biochemistry, Osaka University School of Medicine, Japan.
Fujii
J
J
Seo
H G
HG
Suzuki
K
K
Matsuoka
T
T
Nakamura
M
M
Tatsumi
H
H
Yamasaki
Y
Y
Kamada
T
T
Taniguchi
N
N
D000594:Amino Acid Oxidoreductases / Q000096:biosynthesis*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D006224:Cricetinae; D003513:Cycloheximide / Q000494:pharmacology; D004247:DNA / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D004587:Electrophoresis, Agar Gel; D015870:Gene Expression; D006801:Humans; D007340:Insulinoma; D007375:Interleukin-1 / Q000494:pharmacology*; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D009569:Nitric Oxide / Q000096:biosynthesis / Q000494:pharmacology*; D019001:Nitric Oxide Synthase; D009599:Nitroprusside / Q000494:pharmacology*; D009707:Nucleosomes / Q000187:drug effects / Q000378:metabolism; D010190:Pancreatic Neoplasms; D010396:Penicillamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000494:pharmacology; D026423:S-Nitroso-N-Acetylpenicillamine; D013997:Time Factors; D014407:Tumor Cells, Cultured
D007375:Interleukin-1; D009707:Nucleosomes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D009599:Nitroprusside; D003609:Dactinomycin; D009569:Nitric Oxide; D026423:S-Nitroso-N-Acetylpenicillamine; D004247:DNA; D003513:Cycloheximide; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D010396:Penicillamine