非瑟酮对肝脏缺血再灌注损伤小鼠肝细胞凋亡的影响  

Effect of fisetin on hepatocyte apoptosis in mice with hepatic ischemia reperfusion injury

作　　者：李泽信[1] 王霄[2] 王迎[1] 张永强[3] 狄文玉[4] 王建国[1] LI Zexin;WANG Xiao;WANG Ying;ZHANG Yongqiang;DI Wenyu;WANG Jianguo(Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;School of Pharmacy,Xinxiang Medical University,Xinxiang 453003,Henan Province,China;Department of Anesthesia,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China;Department of Patho-logy,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)

机构地区：[1]新乡医学院第一附属医院肝胆胰外科,河南卫辉453100 [2]新乡医学院药学院,河南新乡453003 [3]新乡医学院第一附属医院麻醉科,河南卫辉453100 [4]新乡医学院第一附属医院病理科,河南卫辉453100

出　　处：《新乡医学院学报》2021年第2期118-123,共6页Journal of Xinxiang Medical University

基　　金：河南省医学科技攻关联合共建项目(编号:LHGJ20200495);新乡医学院第一附属医院青年基金资助项目(编号:QN-2017-B001)。

摘　　要：目的探讨非瑟酮(fisetin)对肝脏缺血再灌注损伤(HIRI)小鼠肝细胞凋亡的影响。方法将18只健康雄性C57BL/6小鼠随机分为假手术(SO)组、HIRI组和HIRI+fisetin组,每组6只。HIRI组和HIRI+fisetin组小鼠夹闭肝左叶和肝中叶的门静脉及肝动脉建立HIRI模型,SO组小鼠仅进行开腹和肝门游离手术。造模前1 h,HIRI+fisetin组小鼠经腹腔注射非瑟酮50 mg·kg^(-1),SO组和HIRI组小鼠经腹腔注射等体积的二甲基亚砜。采用酶联免疫吸附试验检测小鼠血清中丙氨酸转氨酶(ALT)和天冬氨酸转氨酶(AST)水平,Western blot法检测小鼠肝组织中血红素加氧酶-1(HO-1)蛋白表达,末端脱氧核苷酰基转移酶介导性dUTP切口末端标记法检测小鼠肝细胞凋亡。将对数生长期AML-12细胞分为对照组、缺氧复氧(HR)组和HR+fisetin组,对照组AML-12细胞正常培养,HR组和HR+fisetin组AML-12细胞制备HR模型,HR+fisetin组AML-12细胞在HR处理前1 h给予20μmol·L^(-1)的非瑟酮预处理。采用细胞计数试剂盒检测AML-12细胞存活率,流式细胞术检测AML-12细胞凋亡。结果HIRI组和HIRI+fisetin组小鼠血清ALT、AST水平显著高于SO组(P<0.05),HIRI+fisetin组小鼠血清ALT、AST水平显著低于HIRI组(P<0.05)。SO组、HIRI组和HIRI+fisetin组小鼠肝组织中HO-1蛋白相对表达量分别为1.03±0.16、1.77±0.25、3.24±0.72,HIRI组和HIRI+fisetin组小鼠肝组织中HO-1蛋白相对表达量显著高于SO组(P<0.05),HIRI+fisetin组小鼠肝组织中HO-1蛋白相对表达量显著高于HIRI组(P<0.05)。SO组、HIRI组和HIRI+fisetin组小鼠肝细胞凋亡率分别为(4.10±0.14)%、(31.29±1.01)%和(14.41±0.91)%,HIRI组和HIRI+fisetin组小鼠肝细胞凋亡率显著高于SO组(P<0.05),HIRI+fisetin组小鼠肝细胞凋亡率显著低于HIRI组(P<0.05)。对照组、HR组和HR+fisetin组AML-12细胞存活率分别为(98.73±1.56)%、(42.73±3.94)%、(86.43±5.17)%,HR组AML-12细胞存活率显著低于对照组(P<0.05),HR+fisetin组AML-12细胞存活Objective To investigate the effect of fisetin on hepatocyte apoptosis in mice with hepatic ischemia reperfusion injury(HIRI).Methods Eighteen healthy male C57BL/6 mice were randomly divided into sham operation(SO)group,HIRI group and HIRI+fisetin group,with 6 mice in each group.The HIRI model of mice in the HIRI group and HIRI+fisetin group was established by clamping the portal vein and hepatic artery in the left and middle lobe of liver.The mice in the SO group were only performed with laparotomy and porta hepatis dissociation.At one hour before modeling,the mice in the HIRI+fisetin group were treated with fisetin 50 mg·kg^(-1)by intraperitoneal injection,while the mice in the SO group and HIRI group were intraperitoneally injected with the same amount of dimethyl sulfoxide.The levels of serum alanine transarninase(ALT)and aspartate transaminase(AST)were detected by enzyme linked immunosorbent assay.The expression of heme oxygenase-1(HO-1)protein in liver tissues of mice was detected by Western blot.The hepatocyte apoptosis of mice was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling.The AML-12 cells in logarithmic growth phase were divided into control group,hypoxia reoxygenation(HR)group and HR+fisetin group.The AML-12 cells in the control group were cultured normally,while the AML-12 cells in the HR group and HR+fisetin group were used to establish HR model.The AML-12 cells in the HR+fisetin group were pretreated with 20μmol·L^(-1)fisetin at one hour before HR.The survival rate of AML-12 cells was detected by cell counting kit,and apoptosis of AML-12 cells was detected by flow cytometry.Results The levels of serum ALT and AST of mice in the HIRI group and HIRI+fisetin group were significantly higher than those in the SO group(P<0.05).The levels of serum ALT and AST of mice in the HIRI+fisetin group were significantly lower than those in the HIRI group(P<0.05).The relative expression of HO-1 protein in liver tissues of mice in the SO group,HIRI group and HIRI+fisetin group

关 键 词：非瑟酮 缺血再灌注 肝细胞凋亡 血红素加氧酶-1 

分 类 号：R657.3[医药卫生—外科学]