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
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
