水流动力学注射对小鼠肝损伤及肝脏的自然修复作用  

Liver injury induced by hydrodynamic injection and natural repair of liver in mice

作　　者：李胜男[1] 张海英[1] 孙千惠 李艳茹[1] 葛鹤 杜瑜[1] 王琳[1] LI Shengnan;ZHANG Haiying;SUN Qianhui;LI Yanru;GE He;DU Yu;WANG Lin(Department of Pathology,School of Basic Medical Sciences,Jilin University,Key Laboratory of Pathobiology, Ministry of Education,Changchun 130021,China)

机构地区：[1]吉林大学基础医学院病理学系病理生物学教育部重点实验室,吉林长春130021

出　　处：《吉林大学学报（医学版）》2018年第2期275-280,共6页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金资助课题(201115112)

摘　　要：目的:观察水流动力学注射所致肝组织损伤的形态学特点,探讨肝脏修复的过程和机制。方法:25只雌性Balb/c小鼠在3s内经尾静脉注射大体积(1.8~2.0mL)生理盐水。根据注射后时间将小鼠分为30min组、8h组、1d组、3d组和7d组,每组5只鼠。6只未注射雌性Balb/c小鼠作为对照组。小鼠内眦静脉取血,检测血清中丙氨酸氨基转移酶(ALT)水平。利用苏木精-伊红(HE)染色观察各组小鼠肝脏形态学,免疫组织化学染色检测各组小鼠肝细胞中增殖细胞核抗原(PCNA)蛋白表达情况。利用Real-time PCR法检测各组小鼠肝组织中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、表皮生长因子(EGF)、肝细胞生长因子(HGF)、转化生长因子α(TGF-α)、Cyclin D1、血管内皮细胞生长因子(VEGF)、Bax和Bcl-2的mRNA表达水平。结果:与对照组比较,8h组小鼠肝质量/初始体质量比值明显下降(P<0.05),7d组差异无统计学意义(P>0.05)。与对照组比较,1d组小鼠血清中ALT水平明显升高(P<0.01),7d组差异无统计学意义(P>0.05)。显微镜下,与对照组比较,30min组小鼠出现肝细胞肿胀及少量出血,每高倍视野下肝细胞数量明显减少(q=4.760,P<0.05)。与30min组比较,8h组小鼠肿胀的肝细胞数量减少,出血坏死灶扩大,每高倍视野下的肝细胞数和双核细胞数均明显增多(q=7.310,P<0.01;q=7.200,P<0.01)。与8h组比较,1d组小鼠出血坏死灶减少,肝细胞数和双核细胞数均减少(q=4.966,P<0.05;q=6.596,P<0.01),但与对照组比较差异无统计学意义(P>0.05)。3d组小鼠坏死灶基本消失,7d组小鼠肝脏结构与对照组相似。与对照组比较,8h组和1d组小鼠肝细胞PCNA指数均明显升高(t=4.458,P<0.01;t=15.557,P<0.01),7d组差异无统计学意义(P>0.05)。与对照组比较,30 min组小鼠胆管细胞PCNA指数明显升高(t=3.985,P<0.01),7d组差异无统计学意义(P>0.05)。30min组TNF-α、IL-6、EGF和VEGF mRNA表达水平明显高于8h组(q=4.952,P<0.05;q=14.Objective:To observe the morphological features of liver injury induced by hydrodynamic injection,and to explore the procedure and mechanism of liver repair.Methods:Twenty-five female Balb/c mice were intravenously injected with large volume of saline solution(1.8-2.0 mL)in 3 s.The mice were divided into 30 min,8 h,1 d,3 d,and 7 d groups according to the post-injection time.Each group contained 5 mice.Six non-injected mice were used as control group.The blood samples from angular vein of the mice were collected to detect the levels of alanine transaminase(ALT).The morphological features of liver tissue of the mice in various groups were observed with HE staining.The proliferating cell nuclear antigen(PCNA)protein expressions in liver tissue of the mice in various groups were detected by immunohistochemical saining.The expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),epidermal growth factor(EGF),hepatocyte growth factor(HGF),transforming growth factor-α(TGF-α),Cyclin D1,vascular endothelial growth factor(VEGF),Bax and Bcl-2 mRNA were determined by Real-time PCR.Results:Compared with control group,the ratio of liver weight/original body weight in 8 h group was significantly decreased(P<0.05),but there was no significant difference in 7 d group(P>0.05).Compared with control group,the serum ALT level in 1 d group was significantly elevated(P<0.01),but there was no significant difference in 7 d group(P>0.05).Compared with control group,the liver tissue of the mice in 30 min group showed hepatocyte swollen and small hemorrhagic area,and the number of hepatocytes per high power field was significantly declined(q=4.760,P<0.05)under microscope.Compared with 30 min group,the number of swollen cells was decreased in 8 h group,the hemorrhagic and necrotic area enlarged,both of the hepatocyte number and binuclear hepatocyte number per high power field were significantly increased(q=7.310,P<0.01;q=7.200,P<0.01).Compared with 8 h group,the hemorrhagic and necrotic areas in 1 d group were reduced,both

关 键 词：水流动力学注射 肝损伤 肝再生 细胞因子 生长因子 

分 类 号：R361.1[医药卫生—病理学]