Abstract
Objective To investigate the effects of hyperbaric oxygen (HBO) on hypoxia inducible factor-1 (HIF-1),p53 and hepatocyte apoptosis,and the protective effects of hyperbaric oxygen on liver injury with hepatic artery hypoxia.Methods Ninety-six male Sprague-Dawley rats were randomly divided into three groups:control group,hepatic artery ligation (HAL) group and HBO therapy group.All animals underwent laparotomy and skeletonized-liver followed by heparin Ringer' s solution perfusion.HAL was carried out in the HAL group and HBO group.HBO therapy was given in HBO group,twice daily,60 min each time,0.25 MPa 2 h after operation.Blood and liver samples were collected at 1st,3rd,7th,and 14th day after operation.The expression of HIF-1 mRNA in the liver tissue was detected by using reverse transcription-polymerase chain reaction (RT-PCR).The expression of HIF-1 and p53 was detected by using immunohistrochemical staining.Apoptosis of hepatocytes was evaluated by the terminal deoxynucleotidyl transferase mediated dUTP-rhodamine nick end labeling (TUNEL).Serum aspartate aminotransferase (ALT) level was determined and histopathologic changes were observed to evaluate the liver injury.Results The expression of HIF-1 mRNA and protein was decreased significantly in HBO group as compared with that in HAL group,and it reached the peak at 3rd and 7th day [(0.484 ± 0.071)％ vs (0.834 ±0.129) ％,P ＜ 0.01 ; (3.3 ± 3.2) ％ vs (7.8 ± 3.1) ％,P ＜ 0.05].The expression of p53 in HBO group was significantly lower than that in HAL group at 3rd,7th and 14th day post-operation [(17.3 ± 8.6)％vs (29.2 ±8.1)％,P＜0.05; (19.2 ±8.3)％ vs (41.2 ±9.1)％,P＜0.01; (16.0 ±5.6)％ vs (28.0 ± 8.3) ％,P ＜ 0.05].Apoptosis index of hepatocytes at 3rd and 7 th day post-operation was lower in HBO group than in HAL group [(10.2±4.5)％ vs (16.6 ±3.5)％; (11.7 ±6.4)％ vs (21.3±7.4)％,P ＜0.05].Serum ALT level was significantly decreased in HBO group as compared with that in HAL group.There were neovessels in the periphery of the liver and no hepatic cell necrosis in HBO group.Conclusion HBO down-regulates the expression of HIF-1 and P53 mRNA,and inhibits apoptosis of heptocytes,exerting the protective effects against liver hypoxia injury. Key words: Hyperbaric oxygen; Hepatic artery ligation; Hypoxia inducible factor-1 ; p53 ; Apoptosis
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