PMID- 10760090
OWN - NLM
STAT- MEDLINE
DCOM- 20000509
LR  - 20131121
IS  - 0085-2538 (Print)
IS  - 0085-2538 (Linking)
VI  - 57
IP  - 4
DP  - 2000 Apr
TI  - Nitric oxide mediates cyclosporine-induced apoptosis in cultured renal cells.
PG  - 1549-59
AB  - BACKGROUND: The clinical use of cyclosporine (CsA) is limited by its 
      nephrotoxicity. Apoptosis, perhaps instigated by increased nitric oxide synthase 
      (NOS) activity, may play a role in such toxicity. METHODS: Human mesangial cells, 
      human tubular cells, human umbilical vein endothelial cells, or murine 
      endothelial cells were cultured with CsA at final concentrations of 0 to 1000 
      ng/mL for 4 to 24 hours. As inhibitors of apoptosis, 0.01 mol/L 
      L-nitromethylarginine (L-NAME) or 1 microg/mL cycloheximide (CHX) was added, 
      whereas 0.01 mol/L sodium nitroprusside (as a nitric oxide donor) was used as a 
      positive control. Apoptosis was assessed by using TUNEL method and by DNA 
      fragmentation by electrophoresis. In addition, NOS enzymatic activity, Northern 
      blots for inducible NOS (iNOS) mRNA, and immunohistochemically demonstrable iNOS 
      protein were evaluated. RESULTS: Within 12 to 24 hours, CsA significantly 
      increased the fraction (8 to 35%) of apoptotic cells in each cell line, according 
      to the dose. Fragmentation of DNA confirmed apoptosis. L-NAME and CHX inhibited 
      the phenomenon, whereas sodium nitroprusside enhanced it. Each cell line 
      significantly increased NOS activity in response to CsA, an effect blunted by 
      L-NAME and CHX. Neither inhibitor modified the increased iNOS mRNA expression 
      elicited by CsA. Positive staining for both iNOS and p53 proteins was observed in 
      all cell lines incubated with CsA that were inhibited by CHX; L-NAME inhibited 
      only p53 staining. CONCLUSIONS: CsA induces apoptosis in various renal cell 
      lines, and this effect is mediated by the induction of iNOS via p53. These 
      effects may contribute to the acellular fibrosis characteristic of late CsA 
      nephrotoxicity.
FAU - Amore, A
AU  - Amore A
AD  - Nephrology and Dialysis Department, Central Laboratory, Regina Margherita 
      Children's Hospital, Torino, Italy. amorenefro@email.oirmsantanna.piemonte.it
FAU - Emancipator, S N
AU  - Emancipator SN
FAU - Cirina, P
AU  - Cirina P
FAU - Conti, G
AU  - Conti G
FAU - Ricotti, E
AU  - Ricotti E
FAU - Bagheri, N
AU  - Bagheri N
FAU - Coppo, R
AU  - Coppo R
LA  - eng
PT  - Journal Article
PL  - United States
TA  - Kidney Int
JT  - Kidney international
JID - 0323470
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (RNA, Messenger)
RN  - 31C4KY9ESH (Nitric Oxide)
RN  - 83HN0GTJ6D (Cyclosporine)
RN  - EC 1.14.13.39 (Nitric Oxide Synthase)
RN  - V55S2QJN2X (NG-Nitroarginine Methyl Ester)
SB  - IM
MH  - Apoptosis/*physiology
MH  - Cells, Cultured
MH  - Cyclosporine/*pharmacology
MH  - Dose-Response Relationship, Drug
MH  - Enzyme Inhibitors/pharmacology
MH  - Humans
MH  - Immunohistochemistry
MH  - Kidney/cytology/*drug effects/enzymology/*physiology
MH  - NG-Nitroarginine Methyl Ester/pharmacology
MH  - Nitric Oxide/*physiology
MH  - Nitric Oxide Synthase/genetics/metabolism
MH  - RNA, Messenger/metabolism
EDAT- 2000/04/12 09:00
MHDA- 2000/05/16 09:00
CRDT- 2000/04/12 09:00
PHST- 2000/04/12 09:00 [pubmed]
PHST- 2000/05/16 09:00 [medline]
PHST- 2000/04/12 09:00 [entrez]
AID - S0085-2538(15)46904-2 [pii]
AID - 10.1046/j.1523-1755.2000.00999.x [doi]
PST - ppublish
SO  - Kidney Int. 2000 Apr;57(4):1549-59. doi: 10.1046/j.1523-1755.2000.00999.x.