单次剂量照射制备大鼠放射性肝纤维化模型的建立与评价胡宗涛

单次剂量照射制备大鼠放射性肝纤维化模型的建立与评价

胡宗涛1，高世乐1

，秦

峰1

，朱

杰1，黄德武3，董六一

2

(1．安徽医科大学临床学院解放军第105医院肿瘤中心，安徽合肥230031;

2．安徽医科大学药理学教研室，教育部抗炎免疫药理学重点实验室，国家中医药管理局中药药理三级实验室;

3．安徽医科大学省级实验动物中心，安徽合肥230032)

摘要：目的

评价单次30Gy 照射制备大鼠放射性肝纤维化模型的可行性、科学性及实用性。方法清洁SD 雄性大鼠48只，

随机分为照射组（A 组）40只、对照组（B 组）8只。VARIN 21-EX 直线加速器对A 组大鼠右肝进行单次照射（30Gy /次），于照

射后第4、8、12、26周处死，每时相点处死8只，B 组于第4周末处死。观察大鼠活体外观及肝脏的大体标本，检测大鼠血清中

AST 、ALT 活性，ELISA 法测大鼠血清中TGF-β1、HA 、PC-Ⅲ、LN 含量、HE 染色于光镜下观察肝脏组织病理改变、MASSON 染色观察大鼠肝脏组织胶原沉积改变；碱水法检测大鼠肝脏组织中Hyp 含量，免疫组化法测肝脏组织TGF-β1蛋白表达情况。结果

与对照组（B 组）比较，A 组受照射肝组织表现出放射性肝炎及肝纤维化，照射组大鼠血清中AST ，ALT 酶活性和TGF-β1水平自第4周明显升高（P ＜0．05或P ＜0．01），分别于第8周和第12周达到高峰；照射组大鼠血清中HA 、PC-Ⅲ、

LN 含量和肝脏组织中Hyp 含量于照射后第4周显著升高（P ＜0．05或P ＜0．01），一直呈上升趋势，照射后26周值最高；照射组大鼠肝脏组织中TGF-β1蛋白表达于照射后第4周也明显升高（P ＜0．05），

照射后第12周达到高峰，第26周开始下降。光镜下可见肝脏病变呈慢性进行性肝纤维化改变。结论单次剂量照射制备的大鼠放射性肝纤维化模型稳定可靠，动物死亡率较低，可以用于实验研究。

关键词：放射性肝纤维化；大鼠；转化生长因子-β1；模型

Establishment and evaluation on the model of radiation-induced

liver injury of rat caused by once dose radiation

HU Zong-tao ，GAO Shi-le ，QIN Feng ，et al

（Center of Cancer ，the No．105hospital of People ’s Liberation army ，Hefei 230031，China ）

Abstract ：Objective

To evaluate the feasibility ，rationality and practicability about the model of radiation-induced liver injury of rat

caused by single 30Gy dose radiation．Methods 48SD rats were randomly divided into irradiation group （A group ，n =40）and control group （B group ，n =8）．A group was irradiated fractionally by VARIN 21-EX linear acceleratorat at right liver （dose 30Gy once ），eight of which were killed on 4rd 、

8th 、12th and 26th week after radiation．And B group were sacrificed on 4rd weekend．Then we observed the living appearance of rats ，general sample of radiated liver．The blood and liver of all rats were collected for further examinations．The level of Hyp in liver tissue and the serum activities of ALT and AST were detected．The level of TGF-β1，HA ，PC-Ⅲand LN of blood serum

were measured by ELISA ，histological change of livers was observed with hematoxylin-eosin （HE ）staining and Masson＇s trichrome staining by optical microscopy．Meanwhile ，the expression of TGF-beta1protein was detected by immunohistochemistry．Results

Compared with

the control group （B group ），Display of radiating hepatitis and liver fibrosis were observed in radiated liver histology of the A group rats．

The activities of AST ，ALT and the level of TGF-β1in serum of the A group rats showed an increase from 4th week after the first radiation

（P ＜0．05or P ＜0．01），reaching the highest level at 8th and 12th week ，respectively．The content of HA ，PC-Ⅲand LN in blood serum and Hyp in liver tissue of the A group rats have significantly increased from 4th week after the first radiation （P ＜0．05or P ＜0．01）．The expression of TGF-beta1protein in liver tissue of the A group rats has significantly increased from 4th week （P ＜0．05）．Histological change of livers was chronic progressive liver fibrosis by light microscopy．Conclusion The model of radiation-induced liver injury of rat

caused once dose radiation is stable and reliable ，has lower death rate ，conform to the practice of clinical radiation therapy．

Key words ：radiation-induced liver injury ；rat ；TGF-β1；model 基金资助：解放军105医院重点课题（No 2010YZ12）

通信作者：董六一，男，教授，硕士生导师，研究方向：药理学，

E-mail ：dongly061@yahoo．com．cn

由于放疗在腹部肿瘤治疗中日渐广泛的运用及骨髓移植辐射预处理技术的开展，特别是近年来将三维适形、组织间插植内照射等放疗新技术用于肝脏肿瘤治疗，使肝脏局部受到大剂量照射，更重要的是战场核辐射及核设施等意外事故导致的肝脏受到大剂量照射，而肝脏在受到较大剂量照射后，会出现急性放射性肝炎、肝脏功能损害、慢性进行性肝纤维化、

肝硬化甚至个体死亡

［1-2］

。但如何模拟临床放疗实际制作放

射性肝纤维化动物模型，对防治放射性肝纤维化的研究显得

非常重要。本实验模拟临床放射治疗的过程，对大鼠进行单次剂量照射制备放射性肝纤维化模型，通过多种途径评价该模型的可行性、科学性、实用性。1材料

1．1

实验动物清洁级SpragueDawleg （SD ）雄性大鼠，鼠龄

为5 6个月，

体重180 220g ，安徽医科大学实验动物中心提供，生产合格证号：SCXK （皖）：2005-

001。