泽泻汤加味方对盐敏感性HBZY21细胞中AngⅡ-NADPH-ROS信号通路的影响  

Impact of Modified Zexie Decoction on the AngⅡ-NADPH-ROS Signaling Pathway in Salt-Sensitive HBZY21 cells

作　　者：崔海鹏[1] 刘玉玲[1] 刘凯[1] 孙晓旭 王途 赵娟[1] 张树峰[1] CUI Haipeng;LIU Yuling;LIU Kai;SUN Xiaoxu;WANG Tu;ZHAO Juan;ZHANG Shufeng(Chengde Medical College,Chengde,Hebei 067000,China)

机构地区：[1]承德医学院,河北承德067000

出　　处：《湖南中医药大学学报》2019年第3期331-336,共6页Journal of Hunan University of Chinese Medicine

基　　金：河北省高等学校科学技术研究项目(ZD2015126);河北省教育厅优秀青年基金项目(YQ2013005);河北省青年拔尖人才项目(冀组字〔2016〕9号);河北省高校重点学科建设项目资助(冀教高〔2013〕4号)

摘　　要：目的探讨泽泻汤加味方对盐敏感性肾小球系膜细胞中AngⅡ-NADPH-ROS信号通路的作用机制。方法采用高盐和AngⅡ诱导大鼠肾小球系膜细胞的方法建立盐敏感性高血压体外细胞模型,设正常组、模型组、阳性药缬沙坦组、泽泻汤加味方(高、中、低剂量)组。CKK-8法测定细胞活性;RT-qPCR方法检测细胞中Agtr1a、Cyba、NOX4的m RNA表达水平;Western blot方法检测细胞中AT1R、P22、P47、NOX4的蛋白质表达水平;活性氧检测试剂盒检测细胞中ROS水平。结果与正常组比较,模型组细胞中Agtr1a、Cyba、NOX4的mRNA表达水平、AT1R、P22、P47、NOX4蛋白表达水平以及ROS表达水平显著升高(P<0.05)。与模型组比较,缬沙坦组及泽泻汤加味方中剂量组Agtr1a、Cyba、NOX4的mRNA表达水平、AT1R、P22、P47、NOX4蛋白表达水平以及ROS表达水平显著降低(P<0.05)。不同浓度泽泻汤加味方组间,中剂量组对AngⅡ-NADPH-ROS信号通路的下调作用最为明显。结论泽泻汤加味方对盐敏感性高血压肾损害的保护作用可能与抑制AngⅡ-NADPH-ROS信号通路有关。Objective To investigate the effect of modified Zexie Decoction on the angiotensin Ⅱ(AngⅡ)-NADPH-reactive oxygen species (ROS) signaling pathway in salt-sensitive glomerular mesangial cells. Methods An in vitro cell model of salt-sensitive hypertension was established by inducing rat glomerular mesangial cells with high salt and AngⅡ. Cells were divided into normal group,model group,positive valsartan group,and high-,medium-,and low-dose modified Zexie Decoction groups. CKK-8 assay was used to determine the cell viability. RT-PCR was used to determine the mRNA expression levels of Agtr1a,Cyba,and NOX4. Western blot was used to determine the protein expression levels of AT1R,P22,P47,and NOX4 in cells. The ROS levels in cells were measured by a ROS assay kit. Results Compared with the normal group,the model group had significantly higher mRNA levels of Agtr1a,Cyba,NOX4,protein levels of AT1R,P22,P47,and NOX4,and ROS levels (P<0.05). Compared with the model group,the valsartan group and the medium-dose modified Zexie Decoction group had significantly lower mRNA levels of Agtr1a,Cyba,and NOX4,protein levels of AT1R,P22,P47,and NOX4,and ROS levels (P<0.05). Among the three modified Zexie Decoction groups,the medium-dose group had the greatest downregulation of the AngⅡ-NADPH-ROS signaling pathway. Conclusion The protective effect of modified Zexie Decoction against renal injury induced by salt-sensitive hypertension might relate to the inhibition of the AngⅡ-NADPH-ROS signaling pathway.

关 键 词：盐敏感性高血压 肾损伤 泽泻汤 血管紧张素Ⅱ AngⅡ-NADPH-ROS信号通路 

分 类 号：R285.5[医药卫生—中药学]