腹腔和皮下注射CCl4致大鼠肝纤维化2种造模方法的比较及自愈性研究

Comparative and self-healing study of two modeling methods in rat liver fibrosis model induced by intraperitoneal and subcutaneous injection of CCl4

目的:探究腹腔和皮下注射CCl4建立大鼠肝纤维化模型的成模和自愈情况,确定较优的造模方法。方法:40只大鼠随机分成4组[正常对照组、腹腔注射低剂量组、腹腔注射高剂量组(下称“腹低组”“腹高组”)、皮下注射组(下称“皮下组”)]。不同注射方式给予不同剂量CCl4,于4、6、8周末分别处死大鼠。HE、Masson染色观察肝脏病理变化,试剂盒检测AST、ALT、LN和PCⅢ水平,Western blot检测α-SMA蛋白表达。结果:同正常对照组相比,腹低组和腹高组4周末肝组织病理改变明显加重,肝脏系数、纤维化阳性面积、AST、ALT、LN、PCⅢ和α-SMA蛋白表达水平显著升高(P<0.05,P<0.01),2组间无明显差异(P>0.05)。停止注射CCl44周后,相应指标有明显改善(P<0.05,P<0.01),但与正常对照组相比,仍有明显差异(P<0.05,P<0.01)。同正常对照组相比,皮下组4周末组织病理及相关指标无明显差异(P>0.05);6、8周末组织病理及相关指标有明显变化(P<0.01),程度略强于4周末腹高组。结论:2种注射方式均可建立肝纤维化模型,其中腹腔组成模时间优于皮下组;腹腔组高低剂量间成模情况无差异,停止注射CCl44周后存在自愈性。

OBJECTIVE To explore the modeling and self-healing of rat liver fibrosis model respectively induced by intraperitoneal and subcutaneous injection of CCl4,so as to determine the optimal modeling method.METHODS Totally 40 rats were randomly divided into four groups[normal control group,intraperitoneal injection of low and high dose groups(referred to as the intraperitoneal low group and intraperitoneal high group),subcutaneous injection group(referred to as the subcutaneous group)].The rats were given different doses of CCl4by different injection methods,and sacrificed at the end of 4,6 and 8 weeks,respectively.HE and Masson staining were used to observe the pathological changes of liver.AST,ALT,LN and PCⅢlevels were detected by kits.The expression ofα-SMA protein was detected by Western blot.RESULTS Compared with those in the normal control group,the pathological changes of liver tissue were significantly aggravated,and the expression levels of liver coefficient,fibrosis positive area,AST,ALT,LN,PCⅢandα-SMA protein were significantly increased in the intraperitoneal low and high dose groups at the end of 4 weeks(P<0.05,P<0.01),while there was no significant difference between the two groups(P>0.05).Four weeks after the injection of CCl4was stopped,the corresponding indexes showed a significant downward trend(P<0.05,P<0.01),while there was still a significant difference compared with those in the normal control group(P<0.05,P<0.01).Compared with those in the normal control group,there was no significant difference in histopathology and related indexes in the subcutaneous group at the end of 4 weeks(P>0.05);at the end of 6 and 8 weeks,histopathological condition and related indexes changed significantly(P<0.01),and the degree was slightly stronger than that of intraperitoneal high group at the end of 4 weeks.CONCLUSION Two injection methods can be used to establish liver fibrosis model in rats.The model formation time of intraperitoneal group is better than that of subcutaneous group;there in no difference in the model formation between intraperitoneal high and low dose groups,and four weeks after the injection of CCl4is stopped,there is self-healing in the groups.