益肾化瘀方含药血清对NRK52E细胞转分化过程中p-Smad2/3、Smad7表达的影响  

Influence of p-Smad2/3 and Smad7 expression in the process of NRK52E cell transdifferentiation with medicated serum of Yishen Huayu Formula

作　　者：王海亮[1] 赵威 高志卿[3] 邢海燕[3] 张琳琪[3] 

机构地区：[1]河南省中医院,郑州450002 [2]漯河市中医院,漯河462000 [3]河南中医学院第一附属医院,郑州450000

出　　处：《中华中医药杂志》2015年第12期4482-4485,共4页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：河南省基础与前沿技术研究(No.102300410124);郑州市金水区科技攻关重点项目(No.金科[2014]33-40)~~

摘　　要：目的:观察益肾化瘀方含药血清对TGF-β1诱导的体外培养的肾小管上皮细胞(NRK52E)转分化过程中p-Smad2/3、Smad7表达的影响。方法:将体外培养的NRK52E细胞随机分为6组:正常对照组,TGF-β1刺激组,5%、10%、20%益肾化瘀方含药血清组,缬沙坦含药血清组,培养48h后取出。观察各组细胞形态变化,采用免疫细胞化学法检测NRK52E细胞p-Smad2/3、Smad7的表达。结果:NRK52E细胞经TGF-β1刺激后,失去原有的铺路石样形态,拉长增大呈梭形,而与益肾化瘀方含药血清共同作用后,细胞形态有所修复,细胞梭形改变减轻;免疫组化显示与正常对照组比较,TGF-β1刺激组细胞p-Smad2/3核表达明显增强(P<0.01),Smad7表达却减弱(P<0.01)。而各药物干预组与单纯TGF-β1刺激组相比,p-Smad2/3的表达随药物浓度增加呈下降趋势(P<0.01),而Smad7表达的阳性率明显增强(P<0.01)。其中20%益肾化瘀方含药血清组与缬沙坦含药血清组的表达无显著差异。结论:益肾化瘀方含药血清通过抑制Smad2/3的磷酸化,上调Smad7表达,显示出对TGF-β1刺激的NRK52E细胞转分化过程的阻抑作用,并与剂量呈正相关。Objective： To observe the influence of p-Smad2/3 and Smad7 expression in the process of renal tubular epithelial cells（NRK52E） transdifferentiation induced by TGF-β1 with medicated serum of Yishen Huayu Formula in vitro. Methods： NRK52 E cells were randomly divided into 6 groups： Normal control group, the TGF-β1 stimulation group, kidney blood medicated serum levels of 5%, 10% and 20% group, valsartan serum group. after 48 h, Observe each cell morphological changes, using immunocytochemistry method to detect the expression of p-Smad2/3 and Smad7 in NRK52 E cell. Results： after TGF-β1 stimulation, NRK52 E cells lost the original form paving stone samples, has long spindle, and with kidney blood serum medicated interaction, cell morphological repair, spindle cells change to reduce; compared with normal control group by immunocytochemistry, it showed that TGF-β1 stimulate cell p-Smad2/3 nuclear express obviously increased（P〈0.01）, Smad7 expression was reduced（P〈0.01）. And the drug intervention group compared with the pure TGF-β1 stimulation group, the expression of p-Smad2/3 declined with the increase of drug concentration（P〈0.01）, while Smad7 expression positive rate obviously increased（P〈0.01）. there was no significant difference between 20% of medicated serum of Yishenhuayu and valsartan serum group. Conclusion： Medicated serum of Yishen Huayu Formula inhibit phosphorylation of Smad2/3, raising Smad7 expression, inhibitory effect of the process of NRK52 E cell transdifferentiation stimulated by TGF-β1, and was positively related with dose.

关 键 词：益肾化瘀方 肾间质纤维化 大鼠近端肾小管上皮细胞 P-SMAD2/3 SMAD7 

分 类 号：R285[医药卫生—中药学]