Background Hyperglycemia may accelerate liver fibrosis.Currently,there is no effective treatment for liver fibrosis induced by type 2 diabetes.The study aim was to investigate whether RhoA/Rho kinase （ROCK） pathway ...Background Hyperglycemia may accelerate liver fibrosis.Currently,there is no effective treatment for liver fibrosis induced by type 2 diabetes.The study aim was to investigate whether RhoA/Rho kinase （ROCK） pathway is involved in liver fibrosis in the rats with type 2 diabetes and define the protective effects of fasudil on livers.Methods A rat model of type 2 diabetes was established by high fat diet combined with streptozotocin （30 mg/kg,intraperitoneal injection）.Animals were randomly assigned to 3 groups：control rats,untreated diabetic rats that received vehicle and fasudil-treated diabetic rats that received ROCK inhibitor fasudil hydrochloride hydrate （10 mg/kg per day,intraperitoneal injection,for 14 weeks）.The morphological features of liver were observed by HE staining.Accumulation of collagen in livers was determined by Masson staining and the measurement of hydroxyproline.The mRNA expression of transforming growth factor-β1 （TGFβ1）,connective tissue growth factor （CTGF）,type-Ⅰ,and type-Ⅲ procollagen was assessed with real-time polymerase chain reaction.The phosphorylation of myosin phosphatase target subunit-1 （MYPT1）and the protein levels of TGFβ1 and α-smooth muscle actin （α-SMA） were evaluated by Western blotting.Results Compared with control rats,untreated diabetic rats showed higher values of collagen and hydroxyproline in livers （P ＜0.01）,the phosphorylation of MYPT1 and the protein levels of TGFβ1 and α-SMA were increased （P ＜0.01）,and the mRNA expression of TGFβ1,CTGF,type-Ⅰ,and type-Ⅲ procollagen was upregulated （P ＜0.01）; compared with untreated diabetic rats,treatment with fasudil signifcantly reduced values of collagen and hydroxyproline （P ＜0.01）,and decreased the phosphorylation of MYPT1 and the levels of TGFβ1 and α-SMA （P ＜0.01）,concomitant with the downregulation of TGFβ1/CTGF,type-Ⅰ,and type-Ⅲ procollagen mRNA expression （P ＜0.01）.Conclusions Fasudil ameliorates liver fibrosis in rats with type 2 展开更多
目的:通过运用益气活血法的代表方剂补阳还五汤对动脉粥样硬化模型的干预,观察主动脉组织Rho激酶,纤溶酶原激活物抑制物-1(PAI-1)及内皮型一氧化氮合酶(e NOS)mRNA的表达、血脂等指标的变化,揭示该方药抗动脉粥样硬化作用的新靶点和新...目的:通过运用益气活血法的代表方剂补阳还五汤对动脉粥样硬化模型的干预,观察主动脉组织Rho激酶,纤溶酶原激活物抑制物-1(PAI-1)及内皮型一氧化氮合酶(e NOS)mRNA的表达、血脂等指标的变化,揭示该方药抗动脉粥样硬化作用的新靶点和新途径。方法:60只大鼠随机分为正常组,模型组,补阳还五汤低、高剂量组(10,20 g·kg-1),辛伐他汀组(0.6 mg·kg-1),补阳还五汤预防组(10 g·kg-1),除正常组外,其余各组维生素D3加高脂饮食诱导大鼠动脉粥样硬化模型,分别ig给予相应药物,补阳还五汤预防组给药的同时造模,造模成功后干预28 d,检测主动脉Rho激酶,PAl-1及e NOS mRNA表达,血脂水平总胆固醇(TC),甘油三酯(TG),高密度脂蛋白(HDL),低密度脂蛋白(LDL)和氧化型低密度脂蛋白(ox-LDL)。结果:与正常组比较,模型组大鼠Rho激酶,PAl-1 mRNA表达量水平明显升高,e NOS mRNA表达量水平明显降低及血脂TC,TG,LDL,ox-LDL水平明显升高(P<0.01),HDL水平明显降低,均具有明显统计学差异(P<0.01),补阳还五汤低、高剂量治疗组、辛伐他汀组、补阳还五汤预防组明显降低大鼠Rho激酶,PAl-1 mRNA表达量水平,明显升高e NOS mRNA表达量水平,明显降低血脂TC,TG,LDL,ox-LDL水平,明显升高HDL水平,均具有明显统计学差异(P<0.05,P<0.01)。结论:补阳还五汤可以下调Rho激酶,PAl-1 mRNA的表达,同时,上调e NOS mRNA的表达水平、降低血脂,具有抗动脉粥样硬化作用,抑制Rho激酶mRNA,PAl-1 mRNA表达及上调e NOS mRNA表达水平可能是其作用机制之一。展开更多
基金supported by the National Natural Science Foundation of China(Nos.81102444,and81102492)the Central Public Scientific Research Institution Fundamental Project(Nos.2011CHX14,and2013ZD02)the Research Fund for the Doctoral Program of Higher Education of China(No.20091106120033)~~
文摘Background Hyperglycemia may accelerate liver fibrosis.Currently,there is no effective treatment for liver fibrosis induced by type 2 diabetes.The study aim was to investigate whether RhoA/Rho kinase （ROCK） pathway is involved in liver fibrosis in the rats with type 2 diabetes and define the protective effects of fasudil on livers.Methods A rat model of type 2 diabetes was established by high fat diet combined with streptozotocin （30 mg/kg,intraperitoneal injection）.Animals were randomly assigned to 3 groups：control rats,untreated diabetic rats that received vehicle and fasudil-treated diabetic rats that received ROCK inhibitor fasudil hydrochloride hydrate （10 mg/kg per day,intraperitoneal injection,for 14 weeks）.The morphological features of liver were observed by HE staining.Accumulation of collagen in livers was determined by Masson staining and the measurement of hydroxyproline.The mRNA expression of transforming growth factor-β1 （TGFβ1）,connective tissue growth factor （CTGF）,type-Ⅰ,and type-Ⅲ procollagen was assessed with real-time polymerase chain reaction.The phosphorylation of myosin phosphatase target subunit-1 （MYPT1）and the protein levels of TGFβ1 and α-smooth muscle actin （α-SMA） were evaluated by Western blotting.Results Compared with control rats,untreated diabetic rats showed higher values of collagen and hydroxyproline in livers （P ＜0.01）,the phosphorylation of MYPT1 and the protein levels of TGFβ1 and α-SMA were increased （P ＜0.01）,and the mRNA expression of TGFβ1,CTGF,type-Ⅰ,and type-Ⅲ procollagen was upregulated （P ＜0.01）; compared with untreated diabetic rats,treatment with fasudil signifcantly reduced values of collagen and hydroxyproline （P ＜0.01）,and decreased the phosphorylation of MYPT1 and the levels of TGFβ1 and α-SMA （P ＜0.01）,concomitant with the downregulation of TGFβ1/CTGF,type-Ⅰ,and type-Ⅲ procollagen mRNA expression （P ＜0.01）.Conclusions Fasudil ameliorates liver fibrosis in rats with type 2
文摘目的:通过运用益气活血法的代表方剂补阳还五汤对动脉粥样硬化模型的干预,观察主动脉组织Rho激酶,纤溶酶原激活物抑制物-1(PAI-1)及内皮型一氧化氮合酶(e NOS)mRNA的表达、血脂等指标的变化,揭示该方药抗动脉粥样硬化作用的新靶点和新途径。方法:60只大鼠随机分为正常组,模型组,补阳还五汤低、高剂量组(10,20 g·kg-1),辛伐他汀组(0.6 mg·kg-1),补阳还五汤预防组(10 g·kg-1),除正常组外,其余各组维生素D3加高脂饮食诱导大鼠动脉粥样硬化模型,分别ig给予相应药物,补阳还五汤预防组给药的同时造模,造模成功后干预28 d,检测主动脉Rho激酶,PAl-1及e NOS mRNA表达,血脂水平总胆固醇(TC),甘油三酯(TG),高密度脂蛋白(HDL),低密度脂蛋白(LDL)和氧化型低密度脂蛋白(ox-LDL)。结果:与正常组比较,模型组大鼠Rho激酶,PAl-1 mRNA表达量水平明显升高,e NOS mRNA表达量水平明显降低及血脂TC,TG,LDL,ox-LDL水平明显升高(P<0.01),HDL水平明显降低,均具有明显统计学差异(P<0.01),补阳还五汤低、高剂量治疗组、辛伐他汀组、补阳还五汤预防组明显降低大鼠Rho激酶,PAl-1 mRNA表达量水平,明显升高e NOS mRNA表达量水平,明显降低血脂TC,TG,LDL,ox-LDL水平,明显升高HDL水平,均具有明显统计学差异(P<0.05,P<0.01)。结论:补阳还五汤可以下调Rho激酶,PAl-1 mRNA的表达,同时,上调e NOS mRNA的表达水平、降低血脂,具有抗动脉粥样硬化作用,抑制Rho激酶mRNA,PAl-1 mRNA表达及上调e NOS mRNA表达水平可能是其作用机制之一。
