抑毒调肝合剂对四氯化碳复合因素所致肝纤维化大鼠的保护作用

Protective Effect of Yidutiaogan Mixture on Hepatic Fibrosis Induced by Carbon Tetrachloride Complex Factors in Rats

目的观察抑毒调肝合剂对四氯化碳(CCl4)复合因素所致大鼠肝纤维化的干预作用。方法将60只SD大鼠随机分为正常对照组,模型对照组,秋水仙碱组(0.2 mg·kg^(-1)),抑毒调肝合剂小、中、大剂量组(2.7,5.4,10.8 m L·kg^(-1)),每组10只。除正常对照组外,其他各组饮用10%乙醇,皮下注射40%CCl4橄榄油溶液,高脂饮食,制备肝纤维化模型,并灌胃相应药物,每天1次,连续8周。取肝脏行Masson染色,观察肝纤维化程度,采用生化、放射免疫、酶联免疫吸附法检测大鼠血清肝功能和肝纤维化指标,以及肝组织基质金属蛋白酶组织抑制因子-1(TIMP-1)、基质金属蛋白酶-1(MMP-1)、MMP-2、MMP-13、转化生长因子β1(TGF-β1)水平。结果与正常对照组比较,模型对照组大鼠血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、碱性磷酸酶(ALP)、玻璃酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(C-Ⅳ)水平显著升高,白蛋白(ALB)含量显著降低(P<0.01);肝组织TIMP-1、MMP-2、TGF-β1水平显著升高,MMP-1、MMP-13水平明显降低(P<0.01)。与模型对照组比较,抑毒调肝合剂中、大剂量组和秋水仙碱组大鼠血清ALT、AST、HA、LN、PCⅢ、C-Ⅳ水平显著降低,ALB显著升高(P<0.05);肝组织TIMP-1、MMP-2、TGF-β1水平显著降低,MMP-1、MMP-13含量明显升高(P<0.05);与模型对照组比较,抑毒调肝合剂小剂量组上述指标均差异无统计学意义(P>0.05)。结论大、中剂量抑毒调肝合剂具有抗肝纤维化作用,该作用可能与调节TIMPs/MMPs、抑制TGF-β1、减少细胞外基质沉积有关。

ObjectiveTo observe the effect of Yidutiaogan mixture on rat liver fibrosis induced by CCl4complex factors.MethodsSixty SD rats were randomly divided into normal control group,model control group,colchicine group(0.2mg·kg-1),Yidutiaogan mixture low,medium,and high dose groups(2.7,5.4,10.8mL·kg-1)(n=10).Except for normal control group,the other groups were given drinking water containing10%ethanol,subcutaneous injection of40%CCl4in olive oil(3mL·kg-1,twice weekly),and high fat diet for8weeks as the model control group,and they were given relevant medicine intragastrically once a day during modeling.The rats were sacrificed,and the livers were harvested and stained with Masson staining to observe liver fibrosis.Serum levels of ALT,AST,ALP,ALB,HA,LN,PCⅢ,CⅣ,and the levels of TIMP1,MMP1,MMP2,MMP13,and TGFβ1in liver tissue were detected by biochemical assay,radioimmunoassay and ELISA.ResultsCompared with normal control group,serum levels of ALT,AST,ALP,HA,LN,PCⅢ,and CⅣwere significantly increased in model control group,while serum level of ALB was significantly decreased(P<0.01);the levels of TIMP1,MMP2and TGFβ1in liver tissue were significantly increased,while the levels of MMP1and MMP13were significantly decreased(P<0.01).Compared with the model control group,serum levels of ALT,AST,HA,LN,PCⅢ,and CⅣas well as the levels of TIMP1,MMP2,and TGFβ1in liver tissue were all significantly decreased in Yidutiaogan mixture medium dose,high dose,and colchicine groups(P<0.05);serum level of ALB as well as the levels of MMP1and MMP13in liver tissue were significantly increased(P<0.05).However,there was no statistical significance in above indexes in Yidutiaogan mixture low dose group(P>0.05).ConclusionYidutiaogan mixture could be used to prevent experimental hepatic fibrosis through regulating TIMPs/MMPs,suppressing TGFβ1,reducing deposition of extracellular matrix in liver.