丹参水溶性成分丹酚酸A对四氯化碳诱导大鼠肝纤维化的防治作用及机制

Prevention effect and mechanism of salvianolic acid A of water-soluble component from Salvia miltiorrhiza on hepatic fibrosis induced by carbon tetrachloride in rats

目的探讨丹参水溶性成分丹酚酸A对四氯化碳诱导大鼠肝纤维化的防治作用及机制。方法选取成年健康雄性SD大鼠48只,随机分为假手术组、模型组、丹酚酸A组和Wnt通道阻滞剂组(DKK-1),每组各12只。除假手术组外均注射CCl4制作大鼠肝纤维化模型。丹酚酸A组于造模8周后给予丹酚酸A 10 mg/(kg·d)腹腔注射,Wnt通道阻滞剂组(DKK-1)于造模8周后予以DKK-1和丹酚酸A 10 mg/kg腹腔注射,而假手术组和模型组均于造模8周后予以等量生理盐水腹腔注射。给药治疗6周后,处死各组实验动物,收集肝右叶中部两块肝组织进行检测,其中一块利用10%的甲醛进行固定并制备石蜡切片;另一块置于-70℃冰箱中保存备用,用于分子生物学检测。结果经Western blot检测发现,模型组的α-SMA、TIMP-1和CollagenⅠ蛋白水平显著高于假手术组,而丹酚酸A组和Wnt通道阻滞剂组(DKK-1)经相应的治疗之后可以显著下调上述指标的表达水平。模型组的α-SMA、MMP-2、TIMP-1、CollagenⅠ和CollagenⅢmRNA表达水平显著高于假手术组,而丹酚酸A组和Wnt通道阻滞剂组(DKK-1)经相应的治疗之后可以显著下调上述指标的表达水平。模型组的Gsk3β、β-catenin表达水平显著高于假手术组,β-catenin蛋白表达水平显著低于假手术组,而丹酚酸A组和Wnt通道阻滞剂组(DKK-1)经相应的治疗之后可以显著下调Gsk3β、β-catenin的表达水平。结论丹酚酸A可能是通过激活Wnt/β-catenin信号通路来抑制肝纤维化。

Objective To explore the preventive effect and mechanism of salvianolic acid A of water-soluble component from Salvia mihiorrhiza on hepatic fibrosis induced by carbon tetrachloride in rats.Methods 48 adult male SD rats were selected and randomly divided into the sham group,the model group,the salvianolic acid A group,and the Wnt channel blocker group （DKK-1）,with 12 rats in each group.Except the sham group,the CCl4 was injected in other three groups to establish hepatic fibrosis model in rats.In the salvianolic acid A group,10 mg/（kg·d） salvianolic acid A was intraperitoneally injected after 8-week modeling.In the Wnt channel blocker group,the DKK-1 and 10 mg/kg salvianolic acid A were intraperitoneally injected after 8-week modeling.For the sham group and the model group,the same volume of saline was intraperitoneally injected after 8-week modeling.The rats were sacrificed 6 weeks after medications.Two liver tissues from the middle of the right lobe of the liver were collected for test.One of them was fixed with 10% formaldehyde and prepared into paraffin sections.The other one was stored in a refrigerator at -70℃ for molecular biological detection.Results By Western blot detection,the expression of α-SMA,TIMP-1 and Collagen Ⅰ proteins in the model group was significantly higher than that in the sham group,while in the salvianolic acid A group and Wnt channel blocker group,the proteins expression mentioned above could be obviously down-regulated after the corresponding therapy.In the model group,the expression levels of α-SMA,MMP-2,TIMP-1,Collagen Ⅰ and Collagen Ⅲ mRNA were much higher compared with those in the sham group,while in the salvianolic acid A group and the Wnt channel blocker group,these indexes were greatly down-regualted after the related therapy.The Gsk3β and β-catenin expression levels in the model group were much higher than those in the sham group.The β-catenin expression level was extremely higher than that in the sham group. In the salvianolic acid A group and the Wnt channel blocker group,after the corresponding therap,the Gsk3β and β- catenin expression levels could be significantly down-regulated.Conclusion Salvianolic acid A may inhibit hepatic fibrosis by activating Wnt/β-catenin signal pathway.