降压通络方对缺氧诱导的大鼠肾小管上皮细胞TGF-β_(1)/Smad信号通路的影响  

Effect of Jiangya Tongluo Recipe on TGF-β_(1)/Smad Signaling Pathway in Rat Renal Tubular Epithelial Cells Induced by Hypoxia

作　　者：李冰[1] 韩琳 马钰[1] 王淑艳 沈一凡 康鹏飞 张晓宇[1] 郝改梅 秦建国[1] LI Bing;HAN Lin;MA Yu;WANG Shu-yan;SHEN Yi-fan;KANG Peng-fei;ZHANG Xiao-yu;HAO Gai-mei;QIN Jian-guo(Dongfang Hospital Affilicated to Beijing University of Chinese Medicine,Beijing,100078;College of Chinese Medicine,Beijing University of Chinese Medicine,Beijing,102488;Institute of Basic Theory for Chinese Medicine,China Academy of Chinese Medicine Science,Beijing,100700)

机构地区：[1]北京中医药大学东方医院,北京100078 [2]北京中医药大学中医学院,北京102488 [3]中国中医科学院中医基础理论研究所,北京100700

出　　处：《中国中西医结合杂志》2021年第5期589-595,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(No.81173407);北京市中医管理局北京中医药“薪火传承3+3工程”建设项目郭士魁名家研究室项目(No.2011-SZ-A-27)。

摘　　要：目的探讨降压通络方对缺氧诱导的大鼠肾小管上皮(NRK-52E)细胞凋亡的相关分子机制。方法制备降压通络方(1.42 g/mL灌胃大鼠)含药血清和缬沙坦(0.83 mg/mL灌胃大鼠)含药血清,采用缺氧诱导大鼠NRK-52E细胞凋亡模型,分为4组:正常组、模型组、降压通络方组和缬沙坦组。采用CCK-8试剂盒检测细胞活力值,Annexin V-FITC/PI双染法检测细胞凋亡率,ELISA法检测细胞上清液转化生长因子β_(1)(TGF-β_(1))蛋白表达,免疫荧光法检测TGF-βRⅠ、Smad2/3蛋白定位,Western Blot法检测TGF-βRⅠ、Smad2/3蛋白表达。结果TGF-βRⅠ、Smad2/3在NRK-52E细胞上广泛表达。与正常组比较,模型组细胞活力值降低(P<0.01),凋亡率升高(P<0.01),TGF-β_(1)、TGF-βRⅠ、Smad2/3表达升高(P<0.01)。与模型组比较,降压通络方组和缬沙坦组细胞活力值升高(P<0.01),凋亡率降低(P<0.05,P<0.01),TGF-β_(1)、TGF-βRⅠ、Smad2/3表达降低(P<0.01,P<0.05)。与缬沙坦组比较,降压通络方组TGF-β_(1)表达升高(P<0.05),细胞活力值、凋亡率、TGF-βRⅠ和Smad2/3表达比较,差异无统计学意义(P>0.05)。结论降压通络方能抑制缺氧诱导的NRK-52E细胞凋亡,其作用机制可能是通过抑制TGF-β_(1)/TGF-βRⅠ/Samd2/3信号通路而实现。Objective To investigate the molecular mechanism of Jiangya Tongluo Recipe(JYTLR)on apoptosis of rat renal tubular epithelial(NRK-52 E)cells induced by hypoxia.Methods The JYTLR(by gastric infusing drug at the dose of 1.42 g/mL to rats)and valsartan(by gastric infusing drug at the dose of 0.83 mg/mL to rats)containing serums were prepared.The NRK-52 E cells apoptosis model was induced by hypoxia and divided into four groups:normal group,model group,JYTLR group and valsartan group.CCK-8 was used to detect the cell viability,Annexin V-FITC/PI Kit was used to detect the apoptosis rate,ELISA was used to detect the expression of transforming growth factorβ_(1)(TGF-β_(1))protein in the supernatant,immunofluorescence was used to detect the protein localization of TGF-βRⅠand Smad2/3,and Western Blot was used to detect the expression of TGF-βRⅠand Smad2/3.Results TGF-βRⅠand Smad2/3 were widely expressed in NRK-52 E cells.Compared with normal group,the cell viability value of model group decreased(P<0.01),the apoptosis rate increased(P<0.01),TGF-β_(1),TGF-βRⅠ,Smad2/3 expression increased(P<0.01).Compared with model group,the cell viability value of JYTLR group and valsartan group increased(P<0.01),apoptosis rate decreased(P<0.05,P<0.01),the expression of TGF-β_(1),TGF-βRⅠand Smad2/3 decreased(P<0.01,P<0.05).Compared with valsartan group,TGF-β_(1) expression increased in JYTLR group(P<0.05),and there was no significant difference in cell viability,apoptosis rate,TGF-βRⅠand Smad2/3 expression(P>0.05).Conclusion JYTLR can inhibit the apoptosis of NRK-52 E cells induced by hypoxia,and its mechanism may be related to the inhibition of TGF-β_(1)/TGF-βRⅠ/Samd2/3 signaling pathway.

关 键 词：高血压肾损害 降压通络方 缺氧 细胞凋亡 转化生长因子β_(1)/转化生长因子β受体Ⅰ/Smad蛋白2/3 

分 类 号：R285.5[医药卫生—中药学]