低氧诱导因子-1α和p53在肝脏低氧损伤中的作用  

The potential role of hypoxia inducible factor-1β and p53 in liver hypoxia injury

作　　者：杨英祥[1] 邱宝安[1] 夏念信[1] 胡敏[2] 周宁新[3] 高丽杰[3] 黄志强[3] 

机构地区：[1]解放军海军总医院肝胆外科,北京100037 [2]解放军海军总医院病理科,北京100037 [3]解放军总医院肝胆外科研究所,北京100853

出　　处：《感染．炎症．修复》2007年第3期158-161,F0003,共5页Infection Inflammation Repair

摘　　要：目的:探讨低氧诱导因子-1α(HIF-1α)、p53及细胞凋亡在大鼠肝脏低氧损伤中的作用。方法:雄性SD大鼠64只,随机分为对照组和肝动脉结扎组(n=32)。各组均行剖腹、肝脏骨骼化及肝素林格液肝脏灌注,肝动脉结扎组行肝动脉结扎术。术后1、3、7、14d留取肝脏组织及血标本。采用反转录-聚合酶链反应(RT-PCR)法检测肝脏组织HIF-1αmRNA表达水平;免疫组织化学方法检测肝脏组织HIF-1α及p53蛋白表达水平;TUNEL法检测肝细胞凋亡。自动生化分析仪检测血清丙氨酸转氨酶(ALT)活性以评价肝损伤。结果:肝动脉结扎后肝组织HIF-1αmRNA及HIF-1α蛋白较对照组明显增高,分别于结扎后3d和7d达峰值[0.834±0.129比0.372±0.048,(7.8±3.1)%比(2.1±1.7)%,P均<0.01]。肝动脉结扎组肝细胞p53均明显高于对照组(P<0.05或P<0.01),于术后7d达峰值[(41.2±9.1)%比(5.2±4.3)%,P<0.01]。肝动脉结扎组肝细胞凋亡指数均明显高于对照组(P<0.05或P<0.01),于术后7d达峰值[(21.3±7.4)%比(3.7±3.5)%,P<0.01]。肝动脉结扎组ALT水平均明显高于对照组。HIF-1α、p53蛋白表达及凋亡细胞都主要位于肝小叶中央静脉周围。结论:HIF-1αmRNA表达增加、HIF-1α蛋白积聚以及p53促凋亡途径的激活在肝脏低氧损伤中可能发挥重要作用。Objective：To investigate the potential role of hypoxia inducible factor-1α（HIF-1α）, p53 and hepatocyte apoptosis in the liver hypoxia injury. Methods：Sixty-four male Sprague Dawley rats were randomly divided into two groups： control and hepatic artery ligation （HAL） group. All animals underwent laparotomy and liver-skeletonization followed by heparin Ringer＇s solution perfusion. Hepatic artery ligation was performed in the HAL group. Blood and liver samples were collected on days 1, 3, 7, and 14 after operation. Liver tissue HIF-1α mRNA was determined by reverse transcription-polymerase chain reaction （RT-PCR）. The protein expressions of HIF-1α and p53 were dectected by immunohistochemical staining. Liver apoptosis was evaluated by the terminal deoxynucleotidyl transferase mediated dUTP-rhodamine nick end labeling （TUNEL）. Serum aspartate aminotransferase （ALT） levels were determined to evaluate liver injury. Results： Liver HIF-1α mRNA and protein expressions were significantly increased in HAL group compared with the control group, and they reached the peak levels on days 3 and7 [0.834±0.129 vs. 0.372±0.048, （7. 8573.1）% vs. （2.1±1.7）%,P〈0.01] with HIF-1α mRNAgene expression prior to the increase of HIF-1α. The expression of p53 in HAL group was higher than that of the control group, and it peaked on day 7 post operation [（41.2±9.1）% vs. （5.2±4.3）% ,P〈0. 01]. Hypatocyte apoptosis index was lower in the control group than in the HAL group（P〈0. 05 or P〈0. 01）, especially on day 7 post operation[（3. 7±3. 5）% vs. （21.3±7. 4）%]. Serum ALT levels were significantly increased in HAL group than in control group. Conclusion： These results indicate that increase in H IF-1α gene expression, accumulation of HIF-1α in the nuclei and activation of p53 apoptotic pathways might play an important role in the hypoxic injury of the liver.

关 键 词：肝动脉结扎 低氧诱导因子-1Α P53 细胞凋亡 肝动脉血栓形成 

分 类 号：R36[医药卫生—病理学]