高血氨致大鼠急性肝损伤新模型的建立与初步分析  

Hyperammonemia-induced hepatic injury in rats： Characterization of a new animal model

作　　者：余祖江[1] 孙冉[1] 刘晓蕊[1] 闫静雅[1] 高晓娟[1] 家彬[2] 阚全程[2] 

机构地区：[1]郑州大学第一附属医院感染科,450052 [2]郑州大学第一附属医院药剂科,450052

出　　处：《中华肝脏病杂志》2013年第6期467-472,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金（30472031）

摘　　要：目的依次建立大鼠急性肝衰竭模型，D-氨基半乳糖（D-gal）急性肝损伤模型和氯化铵（NH4e1）诱导急性肝损伤模型，探索肝衰竭中高血氨再次诱导肝细胞损伤机制。方法（1）大鼠急性肝衰竭动物模型建立：将雄性SD大鼠36只分为等渗盐水对照组（n=10），12h急性肝衰竭模型组（n=10），24h急性肝衰竭模型组（力=16）；D-gal400rng／kg＋脂多糖50μg／kg混合剂量给予各组大鼠腹腔注射，分别在给药后12h或24h处死各组大鼠。（2）NH4Cl所致肝脏损伤动物模型的建立：将雄性SD大鼠40只分为3组：NH4C1模型组（力=20），D-gal肝脏损伤组（力=10）和等渗盐水对照组。了=10）；NH。C1模型组每8h给予NH。Cl10ml／kg灌胃，急性D-gal肝脏损伤组则于大鼠腹腔每6d注射1次D-gal（800mg／kg），对照组为等渗盐水10ml／kg灌胃，30d后处死各组大鼠。（3）分别检测大鼠血氨，血清丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）；凝血酶原时间活动度比率（PT-A），甲胎蛋白（AFP），Y-谷氨酰转移酶（GGT）；肝脏HE染色后观察病理变化和细胞凋亡指数；统计学分析采用秩和检验。结果在大鼠急性肝衰竭动物模型中，12h后出现转氨酶升高【ALT和AST分别为（1202．51士282．00）U／L和（1560．14±298．98）U／L]，血氨随之升高[（165．9土23．6）μmol／L]，24h检测ALT、AST和血氨分别为（774．40土207．65）U／L、（967．60±121．94）U／L和（143．4-4-18．1）Hmol／L，与等渗盐水对照组比较，尸值均〈0．05。24h后转氨酶和血氨下降，但未发现AFP（除急性肝衰竭24h组外）、GGT变化肝脏病理检查显示各组肝细胞出现片状弥漫性出血性坏死，淤血，并有灶状出血，伴随细胞凋亡指数逐渐上升。但在NH。C1所致急性肝脏损伤中，6h后即检测到血氨升高，ALT和AST同比增加，30d后检测血氨，ALT和AST进一步升高，但AFP，GGT与对�Objective To establish an accurate new rat model ofhyperammonemia-induced liver injury for use in studies of the molecular mechanisms underlying acute liver failure （ALF）. Methods Twentysix Sprague-Dawley rats were administered D-galactosamine （400 mg/kg） and endtoxin （50 μg/kg） viaintraperitoneal injection to induce ALF and sacrificed at 12 h post-injection （ALF-12 group, n = 10） or 24 h post-injection （ALF-24 group, n = 16）. Ten rats adminstered physiological saline served as the control group. In addition, 20 rats were given serial oral administrations of 10% NH4C1 solution （10 ml/kg, every 8 hrs） to establish the hyperammonemia-induced liver injury model; an additional 20 rats were prepared in parallel to serve as the ALF control group （n = 10; D-galactosamine at 800 mg/kg every 6 d for 30 days） and the physiological saline control group （n = 10）. Serum samples were collected from each mouse and used to detect markers of liver function, including alanine aminotransferase （ALT）, aspartate aminotransfease （AST）, alpha-fetal protein （AFP）, and y-glutamyltransferrase （GGT）, as well as blood ammonia （BA） level and prothrombin time activity （PT-A）. Affects on liver histology was assessed by hematoxylin and eosin staining of resected liver tissues, and on apoptosis by TUNEL assay and calculating the apoptotic index （AI）. Results ALF rats showed elevated levels of ALT （1202.51 ＋ 282.00 U/L）, AST （1560.14 ~ 298.98 U/L）, and BA （165.9 ~ 23.6 lamol/L） as early as 6 hrs after model establishment; these levels peaked at 12 hrs after model establishment （ALT： 774.40 -4- 207.65 U/L; AST： 967.60 ＋ 121.94 U/L; BA： 143.4 -4- 18.1 ~rnol/L; P 〈 0.05）. No significant variations were detected in the levels of AFP （except for the ALF-24 group） or GGT. Liver tissues of the ALF-12 and ALF-24 groups showed large or diffuse hemorrhagic necroses with sinusoidal congestion or spotty bleeding, as well as increased AI. Hyperammonemia-induced liver

关 键 词：肝功能衰竭 急性 模型 动物t 高血氨 发病机制 

分 类 号：R575.3[医药卫生—消化系统] R-332[医药卫生—内科学]