γ射线照射联合部分肝切除诱导小鼠肝脏老化  

Liver aging induced by γ-ray irradiation and partial hepatectomy in mice

作　　者：吴勤录 韩子轩 向慎思 万悦 王效芳 董典典 陈慧 于淼 WU Qinlu;HAN Zixuan;XIANG Shensi;WAN Yue;WANG Xiaofang;DONG Diandian;CHEN Hui;YU Miao(School of Basic Medicine,Anhui Medical University,Hefei 230032,China;School of Life Sciences,Hebei University,Baoding,Hebei 071002,China;Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China)

机构地区：[1]安徽医科大学基础医学院,合肥230032 [2]河北大学生命科学院,河北保定071002 [3]军事科学院军事医学研究院,北京100850

出　　处：《军事医学》2024年第10期721-729,共9页Military Medical Sciences

基　　金：国家自然科学基金项目(No.82070596)。

摘　　要：目的鉴于老化在肝细胞移植中的重要作用,建立照射联合肝切除诱导小鼠肝脏老化模型。方法采用60Coγ射线单次照射联合肝切除术造模;造模后检测小鼠存活率、体重、肝指数、肝功,酶联免疫吸附分析(ELISA)及流式细胞术检测血清老化相关分泌因子表达,实时定量PCR(qPCR)检测肝脏组织老化相关分泌因子及端粒酶mRNA水平;Western印迹检测细胞周期依赖性激酶抑制因子1A(CDKN1A)和细胞周期依赖性激酶抑制因子2A(CDKN2A)的表达;ELISA检测肝脏组织中老化相关β-半乳糖苷酶(SA-β-Gal)、脂褐素水平;苏木精-伊红(HE)染色和油红O染色检测肝细胞核大小及脂质积累,称重法检测肝脏中三酰甘油水平;硫代巴比妥酸(TBA)法检测组织丙二醛水平。结果照射联合肝切除后小鼠体重显著降低,肝重体重比并未受到明显影响,转氨酶水平显著升高;SA-β-Gal以及脂褐素水平增加并显著降低端粒酶活性,细胞核明显增大;CDKN1A和CDKN2A的表达增加;老化相关分泌因子的水平显著升高;肝脏脂质沉积及氧化损伤加重。结论成功建立照射联合肝切除诱导小鼠肝脏老化模型。Objective To establish a mouse model of liver aging induced by irradiation combined with hepatectomy.Methods A model was established via single irradiation combined with hepatectomy.The survival rate,body weight,liver index and liver function of the mice were detected.The expressions of senescence-associated secretory phenotype factors in serum were detected with enzyme-linked immunosorbent assay(ELISA)and flow cytometry.The mRNA levels of senescence-associated secretory phenotype factors and telomerase in liver tissue were detected by real-time quantitative PCR(qPCR).The expressions of cyclin dependent kinase inhibitor 1A(CDKN1A)and cyclin dependent kinase inhibitor 2A(CDKN2A)were determined by Western blotting.ELISA was used to calculate senescence-associated β-galactosidase(SA-β-Gal)and lipofuscin levels.Tissue malondialdehyde levels were measured using the thiobarbituric acid(TBA)method.The size of hepatocyte nuclei and lipid accumulation were detected by hematoxylin-eosin(HE)stainingand oil-red-O while triglyceride levels in the liver were studied with the weighing method.Results After irradiation combined with hepatectomy,the body weight of mice was significantly reduced,the liver index was not significantly affected,but the transaminase level was significantly increased.The levels of SA-β-Gal and lipofuscin increased while telomerase activity decreased significantly,and the nucleus size increased.The expressions of cyclin dependent protein kinase inhibitors CDKN1A and CDKN2A increased.The levels of senescence-associated secretory phenotype factors were significantly Hepatic lipid deposition and oxidative damage were aggravated.Conclusion A mouse model of liver aging induced by irradiation combined with hepatectomy has been established.

关 键 词：小鼠 老化 照射 肝切除术 氧化损伤 老化相关分泌表型 

分 类 号：R575[医药卫生—消化系统]