翻白草含药血清对高糖培养肾小管上皮细胞增殖作用及其对RhoA/ROCK信号通路的影响  

Effect of Potentilla purpurea on Proliferation and Rho A/ROCK Signaling Pathway of High Glucose-induced Renal Tubular Epithelial Cells

作　　者：黄雅薇 赵宗江[1] 苗永辉[1] 张新雪[1] 杨冠男[1] 王婷[1] 张紫嫣 

机构地区：[1]北京中医药大学中医学院,北京100029

出　　处：《中国实验方剂学杂志》2017年第17期104-109,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81373831)

摘　　要：目的:探讨翻白草含药血清对高糖培养小鼠肾小管上皮细胞增殖及其对转分化机制的影响。方法:制备各组大鼠含药血清;10%胎牛血清(FBS)-RPMI 1640培养基培养小鼠肾小管上皮细胞,传代培养后按3 000个/孔接种于96孔培养板,分为正常组、高糖组、厄贝沙坦组及翻白草组;加入相对应6%含药血清的高糖RPMI 1640培养基;培养12,24,48,60 h时分别观察各组细胞形态,并应用噻唑蓝(MTT)比色法检测各组吸光度A;高糖培养24 h后应用蛋白免疫印迹法(Western blot)检测各组肾小管上皮细胞小G蛋白(RhoA),Rho蛋白激酶1(ROCK1),α-平滑肌肌动蛋白(α-SMA)和钙黏附蛋白-E(Ecadherin)蛋白表达情况。结果:高糖培养后肾小管上皮细胞由扁平不规则的多角形变为长梭形;加入相应含药血清干预后细胞呈扁平不规则的多角形;与正常组比较,12 h时高糖组肾小管上皮细胞呈明显增殖状态(P<0.05),24,48,60 h时增殖显著(P<0.01);与高糖组比较,12 h时翻白草组能抑制肾小管上皮细胞增殖(P<0.05),24,48,60 h时翻白草组抑制肾小管上皮细胞增殖作用显著增强(P<0.01);与厄贝沙坦组比较,48,60 h翻白草组能抑制肾小管上皮细胞增殖(P<0.05);与模型组比较,翻白草组E-cadherin蛋白表达显著增加,RhoA,ROCK1和α-SMA蛋白表达显著减少(P<0.01)。结论:翻白草含药血清可以抑制高糖培养的肾小管上皮细胞增殖、抑制肾小管上皮细胞转分化,可能与RhoA/ROCK信号通路有关。Objective： To investigate the effect of serum containing Potentilla purpurea on the proliferation and transdifferentiation of renal tubular epithelial cells in mice with high glucose. Method： Serum containing drugs were prepared for various groups. 10% fetal bovine serum （FBS） -roswell park memorial institute 1640 （RPMI 1640） medium was used for culture of renal tubular epithelial ceils in mice and then they were planted on 96 well plate with 3 000 cells per well. The cells were randomly divided into 4 groups： normal group, high glucose group, irbesartan group and P. purpurea group; 6% serum containing corresponding drugs was added to high glucose RPMI 1640 medium; and then the cell morphology was observed in all groups at 12, 24, 48, and 60 h. In addition, absorbance A of all groups was detected by the tetramethylthiazolate （MTT） eolorimetric method. Ras homologous gene family member A （RhoA） , Rho protein kinase 1 （ROCK1） , a-smoothmuscleaetin （a-SMA） and E-cadherin （cadherin-E） protein expression levels were detected by Western blot after 24 hours of high glucose culture. Result ： In high glucose group, the form of renal tubular epithelial cells changed from the fiat irregular polygon into long fusiform; after corresponding drug-containing serum was added, the cells became fiat in irregular polygon. The glucose group of renal tubular epithelial cells showed obvious proliferation condition at 12 h （P 〈 0.05） and extremely significant proliferation at 24, 48, 60 h as compared with normal group （P 〈 0. 01 ）. P. purpurea group could inhibit renal tubular epithelial cell proliferation at 12 h （ P 〈 0.05 ） , and the effect was significantly enhanced at 24, 48, 60 h （P 〈 0.01 ） as compared with high glucose group. P. purpurea group could inhibit renal tubular epithelial cell proliferation at 48, 60 h as compared with irbesartan group （P 〈 0. 05）. In addition, as compared with model group, the expression of E-cadherin protein was increased significantly

关 键 词：翻白草 肾小管上皮细胞 小G蛋白/Rho蛋白激酶1(RhoA/ROCK)信号通路 上皮-间充质转分化 

分 类 号：R285.5[医药卫生—中药学]