Preventive effects of two PAF-antagonists, PMS 536 and PMS 549, on cyclosporin-induced LLC-PK1 oxidative injury.
The present study was undertaken to evaluate the effects of platelet activating factor (PAF) antagonists, PMS 536 and PMS 549, on LLC-PK1 toxicity induced by Cyclosporin A (CsA). The LLC-PK1 cell line was used as an in vitro model. CsA cytotoxicity was determined in relation with ATP content. Alkaline phosphatase and N-acetyl-beta-glucosaminidase activities, which are directly correlated with tubular cell damage, were used as markers for renal injury. CsA alone provoked in the LLC-PK1 cell line a marked decrease in cell viability (55%) and membrane integrity (56%), and a significant increase in AP and NAG activities and in oxidized glutathione level. The ATP decrease and the ADP increase, resulting in a decline of the ATP/ADP ratio, is indicative of an anoxic energy charge. Co-treatment with CsA plus PMS 536 or PMS 549 resulted in a minor decrease in cell viability and in significant membrane integrity recovery. Moreover, the ATP depletion and the increase in ATP metabolites, hypoxanthine and uric acid induced by CsA were strongly prevented by PAF antagonists. In contrast, GSSG level remained high as in CsA-treated cells, but GSH level was in the range of controls. Our results suggest that both PAF antagonists attenuate CsA oxidative injury and prevent energy metabolism disturbances probably by maintaining cell integrity. The lipophilicity of both molecules may be responsible for membrane stabilization and may confer the protective effects observed in energy metabolism. The results obtained with PMS 536 and PMS 549 are indicative of interactions between PAF and CsA in renal injury and suggest the therapeutic potential of these PAF-antagonists against CsA-induced nephrotoxicity.
Massicot
F
F
Laboratoire de Toxicologie (EA 207), Faculté de Pharmacie, Université René-Descartes, Paris, France.
Lamouri
A
A
Martin
C
C
Pham-Huy
C
C
Heymans
F
F
Warnet
J M
JM
Godfroid
J J
JJ
Claude
J R
JR
D000118:Acetylglucosaminidase / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D016572:Cyclosporine / Q000494:pharmacology* / Q000633:toxicity; D005978:Glutathione / Q000378:metabolism; D005979:Glutathione Peroxidase / Q000378:metabolism; D007674:Kidney Diseases / Q000139:chemically induced / Q000517:prevention & control; D018374:LLC-PK1 Cells; D018384:Oxidative Stress / Q000187:drug effects*; D010879:Piperazines / Q000494:pharmacology*; D010972:Platelet Activating Factor / Q000037:antagonists & inhibitors*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology*; D013552:Swine
C105667:PMS 536; C105668:PMS 549; D010879:Piperazines; D010972:Platelet Activating Factor; D010975:Platelet Aggregation Inhibitors; D000244:Adenosine Diphosphate; D016572:Cyclosporine; D000255:Adenosine Triphosphate; D005979:Glutathione Peroxidase; D000469:Alkaline Phosphatase; D000118:Acetylglucosaminidase; D005978:Glutathione