扶脾柔肝方配方颗粒对肝纤维化大鼠肝组织诱导型一氧化氮合酶表达的影响

Effect of Fupi Rougan Formula Granule on Liver Tissue iNOS Expression in Hepatic Fibrosis Rats

目的:观察扶脾柔肝方配方颗粒对肝纤维化大鼠肝组织诱导型一氧化氮合酶(i NOS)表达的影响,并探讨其作用机制。方法:SD大鼠按随机数字表法分为正常组、模型组、秋水仙碱(0.2 mg·kg-1)组、扶正化瘀(0.415 g·kg-1)组、扶脾柔肝方配方颗粒高、中、低剂量(80,40,20 g·kg-1)组,采用四氯化碳复合乙醇诱导大鼠肝纤维化模型,造模8周成功后,分别给予相应药物ig,每日1次,连续4周,正常组、模型组ig等体积生理盐水。第12周末,检测大鼠血清丙氨酸氨基转移酶(ALT),天门冬氨酸氨基酸转移酶(AST),透明质酸(HA);苏木素伊红(HE)染色观察肝组织病理情况;采用实时荧光定量PCR(Realtime PCR)及免疫印迹法(Western blot)检测大鼠肝组织i NOS mRNA和蛋白表达水平。结果:与正常组比较,模型组大鼠血清中ALT,AST,HA含量显著升高(P<0.01);与模型组比较,秋水仙碱组、扶正化瘀组、扶脾柔肝方低、中剂量组血清中的ALT,AST含量均明显降低(P<0.05),高剂量组ALT,AST含量显著降低(P<0.01),秋水仙碱组、扶正化瘀组、扶脾柔肝方中剂量组血清中的HA含量明显降低(P<0.05),扶脾柔肝方高剂量组HA含量显著降低(P<0.01)。与模型组比较,各药物干预组肝组织内纤维化程度均有不同程度减轻(P<0.05)。与正常组比较,模型组大鼠肝组织i NOS mRNA和蛋白表达显著升高(P<0.01);与模型组比较,各药物干预组大鼠肝组织i NOS mRNA和蛋白表达显著降低(P<0.01),其中以扶脾柔肝方高剂量组最明显。结论:扶脾柔肝方配方颗粒下调纤维化肝组织i NOS mRNA和蛋白表达水平,可能是其抗肝纤维化作用机制之一。

Objective： To observe the effect of Fupi Rougan formula granule on liver tissue inducible nitric oxide synthase（ i NOS） expression in hepatic fibrosis rats and explore its action mechanism. Method： SD rats were randomly divided into normal group,model group,colchicine group（ 0. 2 mg·kg- 1）,Fuzheng Huayu group（ 0. 415 g·kg- 1）,Fupi Rougan low-dose（ 20 g·kg- 1） group,Fupi Rougan middle-dose（ 40 g·kg- 1）group,and Fupi Rougan high-dose（ 80 g·kg- 1） group. SD rat models of hepatic fibrosis were induced by carbon tetrachloride composite ethanol（ CCl4）. After 8 weeks,the rats in various groups were given with corresponding drug intervention respectively by intragastric administration,once daily for 4 weeks. The rats in normal group and model group were given with normal saline. The levels of alanine aminotransferase（ ALT）, aspartateaminotransferase（ AST） and hyaluronic acid（ HA） of rat serum were detected at the end of 12 th week. HE staining was used to observe the pathological changes in liver tissue. Fluorescence Real-time PCR and Western blot assay were used to detect mRNA and protein expression levels of i NOS in rat liver tissues. Result： As compared with normal group,the contents of ALT,AST and HA in rat serum were significantly increased in model group（ P 0. 01）. As compared with model group,the contents of ALT and AST in colchicine group,Fuzheng Huayu group,Fupi Rougan low-dose group and Fupi Rougan middle-dose group were decreased（ P 0. 05）,the contents of ALT and AST in Fupi Rougan high-dose group were significantly increased（ P 0. 01）,the contents of HA in rat serum of colchicine group,Fuzheng Huayu group and Fupi Rougan middle-dose group were decreased（ P 0. 05）,the content of HA in Fupi Rougan high-dose group was significantly decreased（ P 0. 01）. As compared with model group,the liver tissues fibrosis was relieved in various degrees in each drug intervention group（ P 0. 05）. As compared with normal group,the mRNA and protein expression levels of the rat liver tissue i NOS were significantly increased in model group（ P 0. 01）. As compared with model group,the mRNA and protein expression levels of the rat liver tissue i NOS were significantly decreased（ P 0. 01） in each drug intervention group,especially in Fupi Rougan high-dose group（ P 0. 01）. Conclusion： The Fupi Rougan formula granule can reduce the mRNA and protein expression levels of the liver fibrosis tissue i NOS. It may be one of the antihepatic fibrosis mechanisms.