缬沙坦对高糖诱导小鼠足细胞损伤的保护机制研究  

Protective mechanism of valsartan on the injured podocyte induced by high glucose

作　　者：叶迅[1] 侯鹏超[1] 洪郁芝[1] 

机构地区：[1]浙江中医药大学附属广兴医院杭州市中医院内分泌代谢科,杭州310007

出　　处：《中国临床药理学杂志》2016年第22期2073-2076,共4页The Chinese Journal of Clinical Pharmacology

基　　金：浙江省中医药科学研究基金计划(A类)基金资助项目(2012ZA097);杭州市科委医疗卫生及重点专科专病科研攻关专项基金资助项目(20130733Q16)

摘　　要：目的观察不同浓度缬沙坦体外干预高糖环境下小鼠足细胞间充质细胞转分化及转化生长因子-β1(TGF-β)/Smad信号通路的调节作用。方法复苏和传代小鼠足细胞,用Rand A 1.0软件完全随机分组法将培养成熟的小鼠足细胞随机分为5 mmol·L^(-1)葡萄糖培养液干预组(A组)和25 mmol·L^(-1)葡萄糖培养液干预组(B组),在25 mmol·L^(-1)葡萄糖培养液中分别加入2×10^(-7)mol·L^(-1)(C组),2×10^(-6)mol·L^(-1)(D组)和2×10^(-5)mol·L^(-1)(E组)缬沙坦。体外培养48 h后,倒置显微镜下观察足细胞形态变化,用实时定量-聚合酶链式反应(RT-PCR)和western-blot技术分别检测各组足细胞肾小球足细胞相关蛋白1(NEPH1)、desmin、TGF-β1和Smad7的mRNA和蛋白表达的变化。结果与A组相比,B组足细胞形态和数量显著变化;C、D、E组足细胞形态和数量均不同程度的接近A组,以E组恢复程度最为明显。A组足细胞NEPH1mRNA为0.75±0.08,NEPH1蛋白为2.30±0.14;Smad7 mRNA为1.97±0.14,Smad7蛋白为1.88±0.16;desmin mRNA为0.54±0.08,desmin蛋白为1.52±0.11;TGF-β1mRNA为0.35±0.08,TGF-β1蛋白为0.22±0.10。B组足细胞NEPH1 mRNA为0.58±0.14,NEPH1蛋白为1.91±0.12;Smad7 mRNA为0.34±0.11,Smad7蛋白为0.34±0.12;desmin mRNA为0.670±0.059,desmin蛋白为2.02±0.17;TGF-β1mRNA为1.69±0.10,TGF-β1蛋白为1.60±0.13,与A组比较,差异有统计学意义(P<0.01)。C组NEPH1 mRNA为0.60±0.15,NEPH1蛋白为1.92±0.13;Smad7 mRNA为0.37±0.16,Smad7蛋白为0.49±0.11;desmin mRNA为0.66±0.13,desmin蛋白为2.01±0.09;TGF-β1mRNA为1.53±0.09,TGF-β1蛋白为1.44±0.11,与A组比较,差异有统计学意义(P<0.01)。D组NEPH1 mRNA为0.63±0.08,NEPH1蛋白为1.95±0.09;Smad7为mRNA 0.66±0.13,Smad7蛋白为0.72±0.12;desmin mRNA为0.62±0.16,desmin为1.96±0.13;TGF-β1mRNA为0.98±0.08,TGF-β1蛋白为0.87±0.11,与B组比较,差异有统计学意义(P<0.01)。E组NEPH1 mRNA为0.72±0.14,NEPH1蛋白为2.13±0.16;Smad7 mRNA为1.41±0.12,Smad7蛋白为1.65±0.12;desmin mRNObjective valsartan on injured pod To explore the possible protective mechanism of ocyte via observing the effect of different doses of valsartan on the epithelial -mesenchymal transition （EMT） and TGF- β/Smad signal pathway in mouse podocyte cultured with highglucose. Methods Matured podocytes were cuhured and randomly divided into 5 mmol . L-1glucose group （group A）, 25 mmol . L-1 glucose group （group B） and 2 × 10^-7 mol . L-1 （group C）, 2 × 10^-6 mol . L-1 （group D） and 2× 10^-5 mol . L-1（ group E） valsartan in terms of RandA 1.0 software after all podocytes were revived and passage cul- tivated. Morphological changes of podocytes were traced with inverted microscope and expression changes of NEPH1, desmin, TGF- 61 and Smad7 were detected by RT- PCR and western blot after 48 h cultured in vitro. Results The morphology and quantity of podocytes in group B significantly changed in comparison with that of group A. While the morphology and quantity of podocytes in group C, D and E were close to group A in varying degrees. Among them, podocytes of group E achieved the most restoration. The podocyte expression of NEPH1 mRNA, NEPH1, Smad7 mRNA, Smad7 ,desmin mRNA, desmin, TGF -β1 mRNA, TGF - β1 in group A were 0. 75 ± 0. 08, 2. 30 ± 0. 14, 1.97±0. 14, 1.88 ±0. 16, 0.54 ±0.08, 1.52 ±0. 11, 0.35 ±0.08, 0.22 ±0. 10. The podocyte expression of NEPH1 mRNA, NEPH1 ,Smad7 mRNA, Smad7 ,desmin mRNA, desmin,TGF- β1 mRNA, TGF- β1 in group B were 0. 58 ±0. 14, 1. 91 ±0. 12, 0. 34 ±0. 11,0. 34 ±0. 12, 0. 67 ±0.06, 2. 02 ±0. 17, 1.69 ±0. 10, 1.60 ±0. 13, had statistical significance with group A （P 〈0. 01 ）. The podocyte expression of NEPH1 mRNA, NEPH1 ,Smad7 mRNA, Smad7, desmin mRNA, desmin, TGF - β1 mRNA, TGF - β1 in group C were 0. 60 ± 0. 15, 1.92 ± 0. 13,0. 37 ± 0. 16, 0. 49 ± 0. 11, 0. 66 ± 0. 13, 2.01 ± 0. 09, 1.53 ± 0. 09, 1.44 ± 0. 11, had statistical significance with group A （P 〈 0. 01 ）. The podoeyte expression of NEPH1 mRNA, NEPH1, Smad7 mRNA, Smad7

关 键 词：缬沙坦 高糖 足细胞 转分化 

分 类 号：R97[医药卫生—药品]