摘要
目的观察高压氧对低氧诱导因子-1（HIF-1）、p．53及细胞凋亡的影响，探讨高压氧对肝动脉性缺氧肝损伤的保护作用。方法将雄性sD大鼠96只随机分为3组：对照组、肝动脉结扎组及高压氧组。各组均行剖腹术、肝门骨骼化及肝素林格氏液肝脏灌注，肝动脉结扎组及高压氧组行肝动脉结扎术。高压氧组术后2h行高压氧治疗，每次吸纯氧60min，2次／d。治疗压力0．25MPa。术后1、3、7、14d留取肝脏及血标本。采用逆转录，聚合酶链反应（RT-PCR）法检测肝脏组织HIF－1 mRNA表达水平；免疫组织化学方法检测肝脏组织HIF－1及p53蛋白表达水平；原位末端转移酶标记（TUNEL）法检测肝细胞凋亡。检测血清丙氨酸转氨酶（ALT）及病理评价肝损伤。结果高压氧组肝脏HIF-1mRNA及蛋白表达较肝动脉结扎组明显下降，分别于治疗后3d和7d达峰值[（0．484±0．071）％比（0．834±0．129）％，P〈0．01；（3．3±3．2）％比（7．8±3．1）％，P〈0．05]；高压氧组术后3、7、14d肝细胞p53表达较肝动脉结扎组均明显降低[（17．3±8．6）％比（29．2±8．1）％，P〈0．05；（19．2．4-8．3）％比（41．2±9，1）％，P〈0．01；（16．0±5．6）％比（28．0±8．3）％，P〈0．05]；高压氧组肝细胞凋亡指数整体水平较肝动脉结扎组低，术后3、7d两组比较差异有统计学意义[（10．2±4．5）％比（16．6±3．5）％；（11．7±6．4）％比（21．3士7．4）％，P〈0．05]；HIF-1、p53蛋白表达及凋亡细胞都主要位于肝小叶中央静脉周围。高压氧组血清ALT含量均较结扎组明显降低（P〈0．01），肝周见新生血管形成。结论高压氧治疗下调HIF-1α mRNA、HIF-1α及p53表达，减少细胞凋亡，对肝脏低氧损伤具有保护作用。
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 1 st, 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 7th 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 hep- tocytes, exerting the protective effects against liver hypoxia injury.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2012年第10期1953-1956,共4页
Chinese Journal of Experimental Surgery
