丹酚酸B对活化的系膜细胞TGF-β_1受体和Smad2表达的影响  

Effect of salvianolic acid B on expression of TGF-β_1 receptors and Smad2 in activated mesangial cells

作　　者：刘煜敏[1] 张悦[1] 陆海英[2] 郝艳鹏[1] 陆敏[1] 

机构地区：[1]上海中医药大学科技实验中心,上海201203 [2]上海中医药大学护理学院,上海201203

出　　处：《中国病理生理杂志》2010年第9期1734-1737,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No30500687);上海市教委重点学科基金资助项目(NoJ50301);上海高校优秀青年教师基金资助项目(Nosyz08044);上海高校创新团队建设项目资助项目

摘　　要：目的:通过观察丹酚酸B对活化的系膜细胞转化生长因子β1(TGF-β1)受体和Sma与Mad的同源基因2(Smad2)分子表达的影响,探讨丹酚酸B拮抗系膜细胞活化及肾纤维化的机制。方法:分离纯化大鼠肾小球系膜细胞,TGF-β1刺激建立系膜细胞活化模型并检测Smad2、Smad7信号分子表达。丹酚酸B(剂量分别为10-6mol/L、10-5mol/L)进行干预,免疫荧光及蛋白印记法观察对α-平滑肌肌动蛋白(α-SMA)的影响,蛋白免疫印迹法检测系膜细胞TGF-β1两型受体(TβRⅠ、TβRⅡ)和Smad2信号分子表达的变化。结果:5μg/L TGF-β1刺激系膜细胞24h可成功制备系膜细胞活化模型,在系膜细胞活化早期即可见Smad2分子的显著磷酸化;10-6mol/L、10-5mol/L丹酚酸B均可明显抑制其活化标志蛋白α-SMA的表达;丹酚酸B干预可致系膜细胞TGF-β1两型受体表达减少,Smad2的磷酸化状态被抑制。结论:丹酚酸B可通过抑制大鼠肾小球系膜细胞TβRⅠ、TβRⅡ表达和信号分子Smad2的磷酸化而拮抗其活化,这可能是丹酚酸B抗肾纤维化的细胞学机制之一。AIM： To study the mechanisms of salvianolic acid B （ Sal B） antagonizing mesangial cell activation and kidney fibrosis through investigating the effect of Sal B on expression of transforming growth factor-β1（ TGF-β1） receptors and Smad2 in TGF-β1-stimulated renal mesangial cell activation. METHODS： Mesangial cells was isolated and purified from rat kidney. TGF-β1 was used to establish rat primary mesangial cell activation model and Smad2,Smad7 protein expression was detected. Sal B （ 10-6 mol/L and 10-5 mol/L） was employed to treat the cells; α-smooth muscle actin（ α-SMA） expression was analyzed by immunofluorescence staining and Western blotting. Mesangial cells were treated with Sal B alone or additional with TGF-β1,and TGF-β1 receptor Ⅰ （ TβRⅠ） ,TGF-β1 receptorⅡ （ TβRⅡ） ,Smad2 phosphorylation and Smad2 protein expression was determined by Western blotting. RESULTS： Cell ular model was established by incubating with 5 μg/L TGF-β1 for 24 h,and in early stage Smad2 was significantly phosphorylated. Sal B （ 10-6 mol/L and 10-5 mol/L） could inhibit α-SMA expression,which was the biomarker of activated mesangial cells. In addition,in Sal B group,the protein expression of TβRⅠand TβRⅡ was significantly down-regulated while Smad2 phosphorylation in mesangial cells was inhibited. CONCLUSION： Sal B inhibits the TGF-β1-Smad pathway,the protein expression of TβRⅠ,TβRⅡ and Smad2 phosphorylation in mesangial cells,which is probably one of the mechanisms of Sal B alleviating kidney fibrosis.

关 键 词：丹酚酸B 系膜细胞 受体 转化生长因子β SMAD2 

分 类 号：R285[医药卫生—中药学]