不同再灌注时间对缺血后大鼠肝组织氧化应激水平的影响  

Effects of different reperfusion times on the level of oxidative stress in rats liver tissue after ischemia

作　　者：陈名阳 胡亚荣 田新雁 陆丽 苏娟 顾伟 Chen Mingyang;Hu Yarong;Tian Xinyan(School of Basic Medicine,Dali University,Yunnan 671000)

机构地区：[1]大理大学基础医学院,云南大理671000 [2]云南省教育厅感染性疾病重点实验室,云南大理671000

出　　处：《中国现代医药杂志》2023年第10期9-14,共6页Modern Medicine Journal of China

基　　金：云南省科技厅地方本科高校(部分)基础研究联合专项(编号:202101BA070001-106)。

摘　　要：目的 探讨不同再灌注时间对缺血后大鼠肝脏损伤程度及氧化应激水平的影响。方法 将42只SD大鼠随机分成假手术对照组(Sham组),缺血组(HI组),缺血再灌注3、6、12、24、72h组(HIR 3、6、12、24、72h组),每组6只大鼠。采用无创血管夹夹闭大鼠肝左外侧叶和中叶的肝动脉、门静脉,阻断约70%入肝血流90min,然后恢复血流再灌注,建立大鼠HIRI模型。Sham组仅进行肝蒂部干扰,HI组不恢复血流再灌注。处死大鼠后,检测血浆中天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)活力,检测肝组织超氧化物歧化酶(SOD)、丙二醛(MDA)、谷胱甘肽(GSH)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、黄嘌呤氧化酶(XOD)、一氧化氮合酶(NOS)、一氧化氮(NO)、层黏蛋白(LN)、透明质酸(HA)的含量;苏木素-伊红(HE)染色观察肝组织病理变化。结果 与Sham组比较,HIR 24h组大鼠肝脏出现了明显的变性和坏死,大鼠肝功能受损。再灌注12h时,血浆中AST、ALT含量显著升高(P<0.01)。SOD、CAT、GSH、GSH-PX等抗氧化酶从再灌注3h起逐渐降低,在12~24h期间变化最明显(P<0.01)。NO和XOD的含量从再灌注6h起开始升高,在12~24h期间变化最明显(P<0.01)。NOS在肝脏缺血和恢复血流再灌注后均出现了降低(P<0.05)。MDA、LN、HA含量随再灌注时间逐渐升高,在72h时达高峰(P<0.01)。结论 再灌注12~24h期间是大鼠肝缺血再灌注损伤的高峰时期,其发生与氧化应激有关。24h后肝损伤虽有所恢复,但肝纤维化变化却在加强。Objective To explore the effect of different reperfusion times on the degree of liver injury and oxidative stress level in rats after ischemia.Methods Forty-two SD rats were randomly divided into sham operation group(Sham group),ischemia group(HI group),ischemia reperfusion groups at 3h,6h,12h,24h and 72h(HIR3,6,12,24,72h group),with 6 rats in each group.The hepatic artery and portal vein of the left lateral lobe and middle lobe of the liver of rats were closed with non-invasive vessel clamp,and about 70%blood flow into the liver was blocked for 90min,and then the blood flow was restored and re-perfusion was established.The Sham group only performed liver pedicle interference,and the ischemia group did not restore blood reperfusion.After the rats were killed,the activity of aspartate aminotransferase(AST)and alanine aminotransferase(ALT)in plasma was detected,the contents of superoxide dismutase(SOD),malondialdehyde(MDA),glutathione(GSH),glutathioneperoxidase(GSH-Px),catalase(CAT),xanthine oxidase(XOD),nitric oxide synthase(NOS),nitric oxide(NO),laminin(LN)and hyaluronic acid(HA)in liver tissue were detected.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of liver tissue.Results Compared with Sham group,ischemia reperfusion injury of rats hepatocytes showed obvious degeneration and necrosis at 24h.The rats liver function was impaired,serum AST and ALT were significantly increased after 12h reperfusion(P<0.01).Antioxidant enzymes such as SOD,CAT,GSH and GSH-Px decreased gradually from 3h after reperfusion,and the changes were most significant from 12~24h(P<0.01).The contents of NO and XOD began to increase from 6h after reperfusion,and the changes were most significant from 12~24h(P<0.01).NOS decreased after liver ischemia and reperfusion(P<0.05).The content of MDA,LN and HA gradually increased with the reperfusion time,reaching a peak at 72h(P<0.01).Conclusion The peak period of hepatic ischemia-reperfusion injury in rats is 12~24h,and its occurrence is related to oxidative stress.Altho

关 键 词：肝缺血再灌注损伤 氧化应激 肝纤维化 

分 类 号：R657.3[医药卫生—外科学] R-332[医药卫生—临床医学]