粒细胞集落刺激因子对大鼠急性肝衰竭肝细胞凋亡的影响  

Effects of granulocyte colony-stimulating factor on hepatocyte apoptosis in acute fiver failure： experiment with rats

作　　者：康文[1] 刘正稳[1] 韩群英[1] 张蕾[1] 雷燕[1] 楼赛[1] 

机构地区：[1]西安交通大学医学院第一附属医院传染科,710061

出　　处：《中华医学杂志》2008年第14期980-984,共5页National Medical Journal of China

基　　金：陕西省科技攻关基金资助项目（2007k14-02）

摘　　要：目的探讨粒细胞集落刺激因子（G-CSF）抑制大鼠急性肝衰竭（ALF）肝细胞凋亡的作用及机制。方法D-氨基半乳糖（D-GaiN）制备SD大鼠急性肝衰竭模型。治疗组皮下注射重组人粒细胞集落刺激因子（rhG-CSF）50μg／kg共3d，安慰剂组皮下注射生理盐水共3d。建模后观察动物生存率，分别于6h、12h、1d、3d取肝脏，流式细胞仪检测肝细胞凋亡率，免疫组化法检测肝脏Bcl-2、半胱氨酸蛋白水解酶（caspase）-3表达，图像分析系统半定量分析。结果治疗组生存率高于安慰剂组（53．3％vs33．3％，P=0．027）。两组肝细胞凋亡率、肝组织Bcl-2、caspase-3表达量均随时间而升高。1d时治疗组肝细胞凋亡率（29％±7％）低于安慰剂组（44％±12％），差异有统计学意义（P=0．026）。治疗组12h、1d时肝组织Bcl-2灰度值均低于安慰剂组，差异有统计学意义（分别为152±37vs161±7，P=0．012；150±12vs159±9，P=0．018），表示Bcl-2表达量高于安慰剂组；治疗组1d、3d时肝组织caspase-3灰度值均高于安慰剂组，差异有统计学意义（分别为189．6±4．6vs169．6±15．7，P=0．000；184．7±4．8vs160．0±5．0，P=0．000），治疗组caspase-3表达量低于安慰剂组。结论肝细胞凋亡在ALF发病过程中发挥了重要作用。G-CSF通过促进肝细胞Bcl-2表达和减少caspaae-3表达，延缓并减少肝细胞凋亡的发生，提高ALF大鼠生存率。Objective To explore the effects of granulocyte colony- stimulating factor （G-CSF） on hepatocyte apoptosis in acute liver failure （ALF） and possible mechanism thereof. Methods One hundred and sixty SD rats underwent intraperitoneal injection of D-galactosamine （ D-GaiN ） 1.4 g/kg so as to establish AFL models and then were randomly divided into 2 equal groups： G-CSF therapy group, injected hypodermically with recombinant human G-CSF 50 μg · kg^-1· d^-1 2 hours after D-GaiN injection for 3 consecutive days, and placebo control group, injected hypodermically with normal saline for 3 consecutive days. Liver samples were collected from 6 rats of each group 6 h, 12 h, 1 day, and 3 days after D-GaiN injection respectively. Another six normal rats were used as normal control group. Hepatocyte apoptosis rate was measured by flow cytometry. Immunohistochemistry was used to detect the expression of Bcl-2 and caspase-3, a proapoptosis protein, in the liver sections. Results The survival rate of the G-CSF therapy group was 53.3%, significantly higher than that of the placebo control group （ 33.3%, P = 0.027 ）. The hepatocyte apoptosis rate and expression rates of Bcl-2 and caspase-3 in the liver sections after D-GaiN injection increased along with time. The hepatocyte apoptosis rate peaked 1 day after the D-GaiN injection in both groups. The maximum hepatocyte apoptosis rate of the G-CSF group was 29% ± 7%, significantly lower than that of the placebo control group （44% ± 12%, P = 0. 026）. The gray scale of Bcl-2 in liver sections at hour 12 of the G-CSF group was 152 ±37, significantly lower than that of the placebo control group （ 161 ± 7, P = 0.012）. and the gray scale of Bcl-2 on day 1 of the G-CSF group was 150 ± 12, significantly lower than that of the placebo control group （ 159 ±9, P =0.018）. The gray scales of caspase-3 on days 1 and 3 of the G-CSF group were 189.6 ±4.6 and 184.7 ±4.8 respectively, both significantly higher than those of the placebo control group （169.6 ±1

关 键 词：粒细胞集落刺激因子 肝功能衰竭 细胞凋亡 流式细胞术 

分 类 号：R686[医药卫生—骨科学]