原代培养大鼠肝细胞在缺氧和复氧条件下的研究：细胞保护剂前列腺素E1、超氧化物歧化酶、别嘌呤醇和维拉帕米的作用

Study of rat hepatocytes in primary culture submitted to hypoxia and reoxygenation: action of the cytoprotectors prostaglandin E1, superoxide dismutase, allopurinol and verapamil.

作者信息

Andrade Dahir Ramos de, Andrade Dahir Ramos de, Santos Sânia Alves Dos

机构信息

Departamento de Moléstias Infecciosas e Parasitárias, Laboratório de Bacteriologia, Faculdade de Medicina, Universidade de São Paulo, São Paulo, Brazil.

出版信息

Arq Gastroenterol. 2009 Oct-Dec;46(4):333-40. doi: 10.1590/s0004-28032009000400016.

DOI:10.1590/s0004-28032009000400016

PMID:20234951

Abstract

CONTEXT

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.

OBJECTIVE

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

METHODS

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.

RESULTS

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.

CONCLUSIONS

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.

摘要

背景

在肝脏疾病中，肝细胞在缺氧和复氧的微环境中暴露于病理状况下的情况非常常见。几种物质对经历缺氧后复氧和单纯缺氧的肝细胞具有细胞保护作用的前景。

目的

我们研究针对遭受缺氧和缺氧+复氧损伤的肝细胞的治疗选择。

方法

在有或没有细胞保护剂的情况下，将大鼠肝细胞原代培养物置于缺氧（2小时）加复氧（2小时）和单纯缺氧（4小时）环境中。通过乳酸脱氢酶释放百分比和细胞活力等功能标准评估肝细胞损伤。在该损伤模型中研究了细胞保护剂前列腺素E1（3μg/mL）、超氧化物歧化酶（80μg/mL）、别嘌呤醇（20μM）和维拉帕米（10⁻⁴M）的作用。