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刘凯,赵娟,刘玉玲,崔海鹏,董雅洁,卢锴锋,孙晓旭,王途,张树峰.泽泻汤加味方通过AT1R通路抑制高盐和AngⅡ诱导HBZY-1中NOX4表达的研究[J].湖南中医药大学学报英文版,2019,39(5):590-595.[Click to copy
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| 泽泻汤加味方通过AT1R通路抑制高盐和AngⅡ诱导HBZY-1中NOX4表达的研究 |
| 刘凯,赵娟,刘玉玲,崔海鹏,董雅洁,卢锴锋,孙晓旭,王途,张树峰 |
| (承德医学院, 河北 承德 067000) |
| 摘要: |
| 目的 探讨泽泻汤加味方通过AT1R通路抑制高盐和AngⅡ诱导的肾小球系膜细胞（HBZY-1）中NOX4表达并初探其作用机制。方法 将HBZY-1细胞随机分为正常组，高盐和AngII诱导模型组，缬沙坦组，泽泻汤加味方中药高、中、低剂量组。造模结束后，缬沙坦组采用缬沙坦（1 μmol/L）刺激，中药组分别用泽泻汤加味方中药高（2 mg/L）、中（1 mg/L）、低（0.5 mg/L）剂量组刺激，正常组正常培养。24 h后收集样本，RT-qPCR方法检测细胞中AT1R、NOX4的mRNA表达水平，MTT法测定细胞活性，Western blot方法检测细胞中AT1R、NOX4的蛋白质表达水平，采用AT1R抑制剂验证中药下调NOX4具体作用通路。结果 与正常组比较，各给药组细胞生长受到明显抑制，RT-qPCR及Western blot结果显示，AT1R、NOX4的mRNA与蛋白表达水平显著降低（P<0.05），中药组与AT1R抑制剂组NOX4表达均下调。结论 泽泻汤加味方可通过下调AT1R、NOX4 mRNA及AT1R、NOX4蛋白水平来保护盐敏感性高血压肾损伤，进而达到延缓肾纤维化的作用，具体作用机制是通过抑制AT1R信号通路进而下调NOX4蛋白表达水平。 |
| 关键词: 盐敏感性高血压 肾纤维化 泽泻汤 血管紧张素Ⅱ AT1R通路 NOX |
| DOI：10.3969/j.issn.1674-070X.2019.05.006 |
| Received:October 31, 2018 |
| 基金项目:河北省高等学校科学技术研究项目（ZD2015126）；河北省青年拔尖人才项目；河北省教育厅优秀青年基金项目（YQ2013005） |
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| Modified Zexie Decoction Inhibits NOX4 Expression in HBZY-1 Induced by High Salt and AngII through AT1R Pathway |
| LIU Kai,ZHAO Juan,LIU Yuling,CUI Haipeng,DONG Yajie,LU Kaifeng,SUN Xiaoxu,WANG Tu,ZHANG Shufeng |
| (Chengde Medical University, Chengde, Hebei 067000, China) |
| Abstract: |
| Objective To investigate the inhibitory effects of modified Zexie Decoction on NOX4 expression in glomerular mesangial cells (HBZY-1) induced by high salt and AngII through AT1R pathway and to explore its mechanism. Methods HBZY-1 was randomly divided into a normal group, a high salt and AngII induced model group, a Valsartan group, a Zexie Decoction modified Chinese herbal medicine high, medium and low dose group. After modeling, the Valsartan group was stimulated by Valsartan (1 1 μmol/L), and the administration group was stimulated by Zexie Decoction with high (2 mg/L), medium (1 mg/L) and low (0.5 mg/L) doses of traditional Chinese medicine respectively. The normal group was cultured normally. After 24 hours, samples were collected. The mRNA expression of AT1R and NOX4 in cells were detected by RT-q PCR, and cell viability was measured by MTT. The protein expression levels of AT1R and NOX4 were detected by Western blot, and specific pathways of NOX4 down-regulation were verified by AT1R inhibitors. Results Compared with the normal group, the growth of cells in each group was inhibited significantly. RT-qPCR and western blot showed that the mRNA expression and the protein expression of AT1R and NOX4 were significantly decreased (P<0.05), while the expression of NOX4 was down-regulated in both Chinese herbal medicine and AT1R inhibited group. Conclusion Modified Zexie Decoction can protect salt-sensitive hypertensive kidney injury by down-regulating AT1R, NOX4, mRNA, AT1R, NOX4 protein levels, and then delay renal fibrosis. The specific mechanism is to down-regulate NOX4 protein expression by inhibiting AT1R signaling pathway. |
| Key words: salt-sensitive hypertension renal fibrosis Zexie Decoction angiotension Ⅱ ATIR pathway NOX4 |
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