四逆散加味抗肝纤维化的作用及机制研究  

Effect and Mechanism of Supplemental Sini San on Hepatic Fibrosis in Rats

作　　者：尚立芝[1] 王付[1] 苗小玲[1] 甘陈菲[1] 张慧娜[1] 

机构地区：[1]河南中医学院经方研究所/方剂学科,郑州450008

出　　处：《中国实验方剂学杂志》2013年第5期207-211,共5页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：河南省科学技术厅科技攻关项目(0424420047);郑州市科技领军人才项目(112PLJRC360)

摘　　要：目的:研究四逆散加味对肝纤维化转化生长因子-β1(TGF-β1)、磷酸化p38丝裂原活化蛋白激酶(p-p38MAPK)、α-平滑肌肌动蛋白(α-SMA)和基质金属蛋白酶(MMP-13)、金属蛋白酶组织抑制因子(TIMP-1)的作用及其相关性,探讨其抗肝纤维化的作用机制。方法:70只Wister大鼠随机分为7组:正常对照组、病理模型组、四逆散组、四逆散加味高、中、低剂量组、四逆散预防组。除正常对照组外,其余各组均采用猪血清ip诱发肝纤维化,0.5 mL/只,2次/周,连续10周,5周后即可形成肝纤维化。预防组于造模同时给药(以四逆散加味7 g.kg-1),各治疗组于造模第6周给药,连续10周。四逆散组4 g.kg-1,四逆散加味高、中、低剂量组(14,7,3.5 g.kg-1)。采用免疫组化S-P法检测肝组织TGF-β1,p-p38MAPK,α-SMA,MMP-13和TIMP-1蛋白阳性染色吸光度(A)。结果:正常组与模型组的TGF-β1依次为[(0.75±0.25)vs(8.67±0.64),P<0.01],p-p38MAPK[(21.71±0.37)vs(80.25±0.53),P<0.01],α-SMA[(3.32±0.64)vs(10.55±0.34),P<0.05]和TIMP-1[(25.57±6.46)vs(200.25±20.36),P<0.01]蛋白表达均显著减弱。四逆散加味中剂量与模型组比较TGF-β1[(5.19±2.33)vs(8.67±0.64),P<0.05],p-p38MAPK[(47.55±0.58)vs(80.25±0.53),P<0.05],α-SMA(6.21±0.78)vs(10.55±0.34),P<0.01)和TIMP-1[(86.02±51.37)vs(200.25±20.36),P<0.01]蛋白表达显著减弱;MMP-13[(109.59±33.21)vs(35.87±6.35),P<0.01]蛋白表达显著增强。结论:四逆散加味有明显的抗肝纤维化作用。其机制可能经TGF-β/p38MAPK信号转导通路抑制肝星状细胞(HSC)活化,使HSC低表达TIMP-1,高表达的MMP-13促进基质降解有关。Objective：To study the effect and mechanism of Supplemental Sini San（SNSJW） on rats liver hepatic fibrosis. Method： The immunohepatic fibrosis model was induced by intraperitoneal injection of porcine serum.in to rats.Seventy Wister rats were divided into seven groups：normol control,model control,SNSJW high-dose group,mediate-dose group and low-dose group,prevention group,Sini San（SNS） group. Except the normol control groups,all the other groups were injected with pig＇s serum without inactivation into abdominal cavity, twice per week and 0.5 mL each time,persisting for 10 weeks. The prevention groups were injected the stomach with the medicines（SNSJW 7 g·kg-1,10 weeks）at the same time when models began to be made, the other treatment groups were dealed with just as the prevention groups 6 weeks later persisting for 4 weeks. Ten rats were randomly taken for decollation in all six groups at the end of 10th week.SNS group 4 g·kg-1,SNSJW high-dose group 14 g·kg-1,mediate-dose group 7 g·kg-1,low-dose group 3.5 g·kg-1.The changes of hepatic fibrosiswere detected respectively.The expression of transforming growth factor-β1（TGF-β1）, p38motogen-activated protein kinase（p38MAPK）,a-smooth muscle actin（α-SMA）,matrixmetalloproteinase-13 （MMP-13）,and tissue inhibitive factor of metalloprote-ase（TIMP-1） in liver tissue were detected by immunohistochemical techniques. Result： Compared with SNSJW mediate-dose group with model group, the expression of TGF-β1, p38MAPK,α-SMA and TIMP-1 protein was lower, but the expression of MMP-13 protein was significantly higher in SNSJW mediate-dose group. Conclusion： Supplemental Sini San has a positive action of treatment on hepatic fibrosis model rats,the mechanism is related with its inhibiting the expression of TGF-β1, TIMP-1 protein and enhancing MMP-13 in rats.

关 键 词：四逆散 肝纤维化 转化生长因子-β1 P38MAPK Α-平滑肌肌动蛋白 基质金属蛋白酶(MMP-13) 

分 类 号：R285.5[医药卫生—中药学]