[1]李泽信,王　霄,王　迎,等.非瑟酮对肝脏缺血再灌注损伤小鼠肝细胞凋亡的影响[J].新乡医学院学报,2021,38(2):118-123.[doi:10.7683/xxyxyxb.2021.02.004]
　LI Zexin,WANG Xiao,WANG Ying,et al.Effect of fisetin on hepatocyte apoptosis in mice with hepatic ischemia reperfusion injury[J].Journal of Xinxiang Medical University,2021,38(2):118-123.[doi:10.7683/xxyxyxb.2021.02.004]
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非瑟酮对肝脏缺血再灌注损伤小鼠肝细胞凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
38
期数:
2021年2
页码:
118-123
栏目:
基础研究
出版日期:
2021-02-05

文章信息/Info

Title:
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
（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）
Author(s):
LI Zexin1; WANG Xiao2; WANG Ying1; ZHANG Yongqiang3; DI Wenyu4; WANG Jianguo1
(1.Department of General Surgery,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China2.School of Pharmacy,Xinxiang Medical University,Xinxiang 453003,Henan Province,China3.Department of Anesthesia,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China4.Department of Patho-logy,the First Affiliated Hospital of Xinxiang Medical University,Weihui 453100,Henan Province,China)
关键词:
非瑟酮; 缺血再灌注; 肝细胞凋亡; 血红素加氧酶-1
Keywords:
fisetinischemia reperfusionhepatocyte apoptosisheme oxygenase-1
分类号:
R657.3
DOI:
10.7683/xxyxyxb.2021.02.004
文献标志码:
A
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20 μmol·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细胞存活率显著高于HR组（P＜0.05）；HR+fisetin组与对照组AML-12细胞存活率比较差异无统计学意义（P＞0.05）。对照组、HR组和HR+fisetin组AML-12细胞凋亡率分别为（6.83±0.21）%、（24.44±0.63）%、（7.63±0.52）%，HR组AML-12细胞凋亡率显著高于对照组（P＜0.05），HR+fisetin 组AML-12细胞凋亡率显著低于HR组（P＜0.05），HR+fisetin组与对照组AML-12细胞凋亡率比较差异无统计学意义（P＞0.05）。结论　非瑟酮预处理能够减轻小鼠HIRI，其机制可能与HO-1蛋白高表达、抑制肝细胞凋亡有关。
Abstract:
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 20 μmol·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 was 1.03±0.16,1.77±0.25 and 3.24±0.72,respectively.The relative expression of HO-1 protein in liver tissues of mice in the HIRI group and HIRI+fisetin group was significantly higher than that in the SO group (P＜0.05).The relative expression of HO-1 protein in liver tissues of mice in the HIRI+fisetin group was significantly higher than that in the HIRI group (P＜0.05).The hepatocyte apoptosis rate of mice in the SO group,HIRI group and HIRI+fisetin group was (4.10±0.14)%,(31.29±1.01)% and (14.41±0.91)%,respectively.The hepatocyte apoptosis rate of mice in the HIRI group and HIRI+fisetin group was significantly higher than that in the SO group (P＜0.05),and the hepatocyte apoptosis rate of mice in the HIRI+fisetin group was significantly lower than that in the HIRI group (P＜0.05).The survival rate of AML-12 cells in the control group,HR group and HR+fisetin group was (98.73±1.56)%,(42.73±3.94)% and (86.43±5.17)%,respectively.The survival rate of AML-12 cells in the HR group was significantly lower than that in the control group (P＜0.05),and the survival rate of AML-12 cells in the HR+fisetin group was significantly higher than that in the HR group (P＜0.05),but there was no significant difference in the survival rate of AML-12 cells between the HR+fisetin group and the control group (P>0.05).The apoptosis rate of AML-12 cells in the control group,HR group and HR+fisetin group was (6.83±0.21)%,(24.44±0.63)% and (7.63±0.52)%,respectively.The apoptosis rate of AML-12 cells in the HR group was significantly higher than that in the control group (P＜0.05),and the apoptosis rate of AML-12 cells in the HR+fisetin group was significantly lower than that in the HR group (P＜0.05),but there was no significant difference in the apoptosis rate of AML-12 cells between the HR+fisetin group and the control group (P>0.05).Conclusion　Fisetin pretreatment can alleviate HIRI in mice,and the mechanism may be related to HO-1 protein overexpression and the inhibition of hepatocyte apoptosis.

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更新日期/Last Update: 2021-02-05