Study of rat hepatocytes in primary culture submitted to hypoxia and reoxygenation: action of the cytoprotectors prostaglandin E1, superoxide dismutase, allopurinol and verapamil.
Exposure of hepatocytes to pathological conditions in a microenvironment of hypoxia and reoxygenation is very frequent in hepatic diseases. Several substances present perspectives for cytoprotective action on hepatocyte submitted to reoxygenation after hypoxia and simple hypoxia.



We research therapeutic options for hepatocytes submitted to hypoxia and hypoxia + reoxygenation injury.



Primary culture of rat hepatocytes was submitted to hypoxia (2 hours) plus reoxygenation (2 hours) and simple hypoxia (4 hours) in the presence or the absence of cytoprotectors. The hepatocyte lesion was evaluated by functional criteria through percentage of lactate dehydrogenase released and cell viability. The effects of the cytoprotectors prostaglandin E1 3 etag/mL, superoxide dismutase 80 microg/mL, allopurinol 20 microM and verapamil 10-4 M were studied in this model of injury.



Reoxygenation after hypoxia induced more significant lesion in cultured hepatocytes compared to simple hypoxia, detected by analysis of functional criteria. There was a significant reduction of percentage of lactate dehydrogenase released and a significant increase of percentage of cell viability in the hypoxia + reoxygenation + cytoprotectors groups compared to hypoxia + reoxygenation groups. Prostaglandin E1, superoxide dismutase and verapamil also protected the group submitted to simple hypoxia, when evaluated by functional criteria.



We conclude that reoxygenation after hypoxia significantly increased the lesion of cultured rat hepatocytes when compared to simple hypoxia. Prostaglandin E1, superoxide dismutase, allopurinol and verapamil acted as cytoprotectors to the rat cultured hepatocytes submitted to hypoxia + reoxygenation in vitro. The substances prostaglandin E1, superoxide dismutase and verapamil protected hepatocytes submitted to simple hypoxia on the basis of all the criteria studied in this experimental model.
Departamento de Moléstias Infecciosas e Parasitárias, Laboratório de Bacteriologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil. dahira@uol.com.br
Dahir Ramos de
DR
Andrade
Dahir Ramos de
DR
Andrade
Sânia Alves Dos
SA
Santos
D000493:Allopurinol / Q000494:pharmacology; D000527:Alprostadil / Q000494:pharmacology; D000818:Animals; D015687:Cell Hypoxia / Q000187:drug effects*; D002478:Cells, Cultured; D019610:Cytoprotection / Q000187:drug effects*; D005260:Female; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000502:physiology; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D010100:Oxygen / Q000008:administration & dosage*; D051381:Rats; D013482:Superoxide Dismutase / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D000493:Allopurinol; D014700:Verapamil; D007770:L-Lactate Dehydrogenase; D013482:Superoxide Dismutase; D000527:Alprostadil; D010100:Oxygen