转基因HBV小鼠急性肝损伤的生物学特性研究

魏琴1,2， 姜涛2， 苗娜3， 张春2， 李志强2， 寿玺2， 马嵋2， 丁剑冰1

(新疆医科大学1基础医学院免疫学教研室， 乌鲁木齐830011；2第一附属医院新疆医学模型动物研究重点实验室；

3第一附属医院病理科， 乌鲁木齐830054)

摘要:目的研究四氯化碳(CCL4)诱导转基因乙型肝炎病毒(HBV)小鼠急性肝损伤的生物学特性的改变。方法选取32只健康雄性C57BL/6小鼠作为空白对照组，实验组：选用32只健康雄性C57BL/6小鼠作为CCL4干预组，32只健康雄性HBsAg表达阳性C57BL/6-HBV转基因小鼠为HBV+CCL4干预组。空白对照组按0.5 mL/kg腹腔注射橄榄油溶液，于0、24、48、72 h 4个时间点取标本。实验组每组各留8只作为空白对照，均按0.5 mL/kg腹腔注射橄榄油溶液，0 h处死；其余24只各自随机分组，一次性给予0.5 mL/kg腹腔注射10%CCL4橄榄油溶液，分别于24、48、72 h每组随机挑选8只小鼠，采集标本，全血用于计数白细胞(WBC)、红细胞(RBC)、血小板(PLT)，取血清测定天冬氨酸转氨酶(AST)、丙氨酸转氨酶(ALT)、碱性磷酸酶(ALP)、总蛋白(TP)、白蛋白(ALB)。结果与空白对照组比较，CCL4干预组24 h AST、ALT明显升高，差异有统计学意义(P<0.01)；HBV+CCL4干预组与CCL4干预组相比，AST、ALT水平在损伤24 h达到峰值，且差异有统计学意义(P<0.01)； CCL4干预组和HBV+CCL4干预组24 h时WBC均呈现下降，且两组差异有统计学意义(P<0.05)；HBV+CCL4干预组RBC在24、48、72 h均降低，与0 h比较差异有统计学意义(P<0.05)；HBV+CCL4干预组PLT 在48、72 h均降低，与0 h比较差异有统计学意义(P<0.05)。HBV转基因小鼠在CCL4诱导下24 h AST、ALT显著升高，WBC、RBC、PLT均降低；ALP、TP、ALB无明显差异，这些指标的改变符合急性肝损伤模型的标准。结论成功建立HBV转基因小鼠急性肝损伤模型，为进一步研究急性肝损伤的发病机制及其他变化提供了可靠的实验平台。

关键词：HBV转基因小鼠； 四氯化碳； 急性肝损伤；

中图分类号：R392文献标识码：A

doi:10.3969/j.issn.1009-5551.2015.04.013

[收稿日期：2014-12-04]

Study on the biological characteristics of HBV transgenic mice with acute liver injury

WEI Qin1,2， JiANG Tao2， MIAO Na3， ZHANG Chun2， LI Zhiqiang2， SHOU Xi2，

MA Mei2， DING Jianbing1

(1DepartmentofImmunology，BasicMedicalCollege，XinjiangMedicalUniversity,

Urumqi830011，China;2XinjiangKeyLaboratoryofMedicalAnimalModelResearch;

3DepartmentofPathology,TheFirstAffiliatedHospital,XinjiangMedicalUniversity,Urumqi830054,China)

Abstract:ObjectiveTo study the biological characteristics of acute liver injury in mice induced by transgenic hepatitis B virus (HBV) by carbon tetrachloride (CCL4). Methods32 male C57BL/6 mice were the control group; 32 male C57BL/6 mice were the CCL4 group; 32 male transgenic C57BL/6 mice with HBsAg positive expression were HBV+CCL4 group. The control group was injected intraperitoneally by 0.5 mL/kg olive oil and taken specimens at 0, 24, 48, 72 h. 8 mice in two model groups were taken as control and given 0.5 mL/kg intraperitoneal injection of olive oil solution, killed at 0 h. The other 24 mice were grouped randomly and given 0.5 mL/kg 10% CCL4 olive oil solution once to produce acute liver injury model, 8 mice among which were randomly selected for specimens at 24, 48, 72 h to detect whole blood leukocyte count (WBC), red blood cell (RBC), platelets (PLT), measured in serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), total protein (TP), albumin (ALB). ResultsCompared with the control group, AST and ALT in CCL4 group peaked at 24 h, with significant difference (P<0.01)； compared with the CCL4 group, AST and ALT in HBV+CCL4 group peaked at 24 h, with a significant difference (P<0.01)；WBC decreased in model groups, with a significant difference (P<0.05)；Compared with 0 h in HBV+CCL4 group; RB C in HBV+CCL4 group decreased at 24, 48, 72 h, with a significant difference (P<0.05)；Compared with 0 h in HBV+CCL4 group，PLT decreased at 48， 72 h, with a significant difference (P<0.05). At 24 h AST and ALT peaked; WBC, RBC, PLT decreased in HBV transgenic mice induced by CCL4; ALP, TP, ALB had no significant difference. All the changes were in accordance with the standard indicators of acute liver injury. Conclu-sion58The establishment of acute liver injury model in HBV transgenic mice was of feasibility and reliability for further research on pathogenesis of acute liver injury.

Key words: HBV transgenic mice; carbon tetrachloride; acute liver injury

乙型肝炎病毒(HBV)的持续存在一直严重影响着人类的肝脏功能，目前全球约有4亿人是HBV携带者，全世界每年因HBV感染及其后遗症而死亡的人数为100万～200万[1]。急性肝损伤又是各种严重肝病发生、发展的始动环节，如果不能得到及时有效的治疗，很快就可转变为慢性肝损伤，甚至是肝纤维化、肝硬化乃至肝癌。因此，肝损伤已经成为严重危害人类健康的重大疾病之一。……