血府逐瘀汤对刀豆蛋白A诱导小鼠肝纤维化的防治作用

Preventive and Therapeutic Effects of “Xuefu Zhuyu Decoction”on Liver Fibrosis Induced by Concanavalin in Mice

目的:探索血府逐瘀汤对刀豆蛋白A(Con A)诱导小鼠肝纤维化的防治作用。方法:将Balb/c雄性小鼠,随机分为正常组、模型组及治疗组。模型组与治疗组按照12.5 mg/kg小鼠体质量予以尾静脉注射Con A,正常组予注射相应剂量生理盐水,1次/周,共10周。治疗组于造模同时予血府逐瘀汤药液10 ml/kg小鼠体质量灌胃,正常组及模型组予以等量饮用水,1次/d。10周后,处死各组小鼠,全自动生化仪检测血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)和白蛋白(Alb);HE和天狼星红染色观察肝组织炎症和胶原沉积;碱水解法检测肝组织羟脯氨酸(Hyp)含量;RT-PCR法检测肝组织α-平滑肌肌动蛋白(α-SMA)、转化生长因子β1(TGF-β1)及Ⅰ、Ⅲ、Ⅳ型胶原(Col I、Ⅲ、Ⅳ)表达。结果:模型组小鼠ALT和AST活性均较正常组明显升高(P<0.05);肝组织HE染色可见肝细胞坏死和大量炎性细胞浸润;天狼猩红染色可见大量胶原沉积和假小叶形成;肝组织α-SMA、TGF-β1及ColⅠ、Ⅲ、ⅣmRNA表达较正常组显著升高(P<0.01)。与模型组相比,治疗组ALT、AST水平明显降低(P<0.05);肝脏炎症和胶原沉积明显改善;α-SMA、TGF-β1及ColⅠ、Ⅲ、ⅣmRNA表达显著下调(P<0.05,P<0.01)。结论:血府逐瘀汤可通过减轻肝脏炎症和抑制肝星状细胞(HSC)活化减少胶原合成而改善Con A诱导的小鼠肝纤维化。

Objective: To investigate the preventive and therapeutic effects of'Xuefu Zhuyu Decoction'on liver fibrosis induced by Con A in mice. Methods: Balb / c male mice were randomly divided into three groups: normal group,model group and treatment group. Mice in the model group and treatment group were i. v. injected Con A at a dose of 12. 5 mg / kg,and mice in the normal group were i. v. injected with normal saline,once a week for ten weeks. When modeling,mice in the treatment group were treated with'Xuefu Zhuyu Decoction'at a dose of 10 ml / kg once a day. Correspondingly,mice in the normal group and model group were treated with water. Ten weeks later,mice were sacrificed. The serum and tissue samples were collect to detect the levels of ALT,AST,Alb; liver inflammation and collagen deposition were observed by HE and Sirius-red staining; content of hydroxyproline was detected by hydrolysis method; the expression of α-smooth muscle actin( α-SMA),transforming growth factor-β1( TGF-β1) and collagen type Ⅰ,Ⅲ and Ⅳ were detected by real-time PCR. Results: Compared to the normal group,the levels of ALT and AST in the model group were significantly increased( P < 0. 05); in the model group,the necrosis of liver cells and infiltration of inflammatory cells can be observed in HE staining,meanwhile collagen deposition and pseudolobule can be seen in Sirius-red staining; the expression of α-SMA,TGF-β1,collagen type Ⅰ,Ⅲ andⅣ mRNA were significantly increased( P < 0. 01). Compared with the model group,the serum level of ALT and AST in the treatment group were reduced markedly( P < 0. 05); hepatic inflammation and the deposition of collagen were improved;expression of α-SMA,TGF- β1,collagen type Ⅰ,Ⅲ and Ⅳ mRNA were down-regulated( P < 0. 05,P < 0. 01).Conclusion: 'Xuefu Zhuyu Decoction'can protect liver tissue against Con A-induced liver fibrosis by alleviating inflammatory response,inhibiting the activation of hepatic stellate cells( HSC) as well as down-regulating the expression of collagens.