黄芪甲苷联合大黄素通过Smads通路抑制TGF-β1诱导的足细胞凋亡  

Astragaloside Combined with Emodin Inhibiting Mice Podocyte Apoptosis Induced by TGF-β_1 via Smad Signal Pathway

作　　者：王宇晖[1] 童孟立[1] 杨汝春[1] 

机构地区：[1]杭州市中医院肾内科,浙江杭州310007

出　　处：《中华中医药学刊》2014年第5期1089-1093,共5页Chinese Archives of Traditional Chinese Medicine

基　　金：浙江省中医药管理局项目(2006C108)

摘　　要：目的：探讨黄芪甲苷联合大黄素是否通过Smads通路抑制转化生长因子-β1（TGF—β1）诱导小鼠足细胞株凋亡。方法：①体外培养小鼠足细胞株。②应用不同浓度TGF—β1（10^-1～10^4ng／L）诱导小鼠足细胞凋亡。③TGF—β1不同刺激时间（12、24、48h）诱导小鼠足细胞凋亡。④黄芪甲苷、大黄素联用抑制TGF—β1诱导小鼠足细胞凋亡，实验分为正常组、TGF—β1组、黄芪甲苷＋TGF—β1组（黄芪甲苷组）、大黄素＋TGF—β1组（大黄素组）、黄芪甲苷＋大黄素＋TGF—β1组（黄芪甲苷＋大黄素组）。⑤采用流式细胞仪检测AnnexinV、Caspase3。⑥实时定量PCR及Western印迹法检测Smad2、Smad3mRNA及蛋白质表达。结果：①随着TGF—β1浓度增加（10^-1～10^4ng／L）、刺激时间增加，足细胞凋亡比率增多（P〈0．05），足细胞Smad2、Smad3mRNA表达增加。TGF—β1 10^3ng／L为最佳诱导小鼠足细胞凋亡浓度，24h为最佳诱导凋亡时间。（2）TGF—β1组与正常组相比Caspase3阳性细胞比率增加（P〈0．01）；Smad2、Smad3蛋白质表达明显增加（P〈0．01）。③黄芪甲苷＋大黄素组与TGF—β1组、黄芪甲苷组、大黄素组比较：足细胞凋亡比率明显减少（P〈0．05）；Caspase3阳性细胞比率减少（P〈0．05）；Smad2、Smad3蛋白质表达减少（P〈0．05）。④黄芪甲苷＋大黄素组较TGF—β1组Smad2、Smad3mRNA表达减少。结论：（1）TGF—β1能通过Smads通路诱导小鼠足细胞凋亡，呈浓度及时间依赖性。②黄芪甲苷联合大黄素能通过Smads通路阻断足细胞凋亡，其机制可能与减少TGF—β1的表达，阻止其激活Smads蛋白，从而阻止Caspase家族诱导的足细胞凋亡有关。Objective ：To investigate the effect of Astragaloside combined with emodin on mice podoctye apoptosis and signal pathway induced by TGF - lSl. Methods ： （1）Mice podocyte strains were divided as following： control group, TGF - β1 （ 10-1 - 10^4 ng/L） groups cultured for 24 hours. （2）TGF -β1 10^3 ng/L treated groups were culture for 12,24,48 hours individually. （3）TGF - β1 treated groups with Astragaloside, emodin or Astragaloside combined with emodin respectively. Annexin V and Caspase3 were measured by flow cytometry. （4）Smad2 and Smad3 mRNA and protein were detected by Real time PCR and Western blotting. Results ： （1）The ratio of apoptotic podocytes and Smad2, Smad3 mRNA expressions were increased after treated with TGF - β1 compared with those of control and correlated with TGF -β1concentration and cultured time （P 〈0.05）. （2）TGF - β1 （103ng/L） and 24 hours treated time were the best way for podocyte apoptosis. （3）The ratio of Caspase3 positive podocytes was increased after treated with TGF - β1 （103ng/L,24h） compared with con- trol （ P 〈 0.05 ）. （4）The ratio of apoptotic podocytes and Smad2, Smad3 and protein level were decreased after treated with atragaloside combined with emodin compared to TGF-β1 ,Astragaloside and emodin groups respectively. （5）The ratio of Smad2 and Smad3 mRNA expressions was decreased after treated with Astragaloside combined with emodin compared to that of TGF- β1 group. Conclusion ：TGF- β1 can enhance the apoptosis of mice podocyte time -dependently and dose - dependently by Smad signal pathway. Astragaloside combined with emodin can reduce the TGF - β1 expression and activa- tion of Smad protein and inhibit mice podocyte apoptosis induced by Caspase.

关 键 词：黄芪甲苷联合大黄素 SMADS 通路 TGF—β 足细胞凋亡 

分 类 号：R258.8[医药卫生—中医内科学]