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阿利克仑对AGT-REN双转基因高血压小鼠肾损伤的保护作用

杨晓玲^1,2, 陈妍妍³, 赵华¹, 张博洋¹, 张晓伏¹, 李晓洁¹, 杨秀红^1,2,*

¹华北理工大学基础医学院，唐山 063000；²河北省慢性疾病基础医学重点实验室，唐山063000；³中直机关医院，北京 100091

摘要

本文旨在研究肾素抑制剂阿利克仑对人血管紧张素原-肾素(angiotensinogen-renin, AGT-REN)双转基因高血压(double transgenic hypertension, dTH)小鼠肾损伤的影响，并探讨其可能的机制。将dTH小鼠分为高血压组(hypertension, HT组)和阿利克仑干预组(HT+Aliskiren组)；野生C57BL/6小鼠作为对照组(wild-type, WT组)。HT+Aliskiren组小鼠皮下渗透阿利克仑(20 mg/kg) 28 d，收集小鼠血压数据；通过HE、Masson、PAS染色观察肾脏组织结构损伤和胶原沉积情况；透射电子显微镜观察肾脏超微结构；考马斯亮蓝染色法和生化分析仪检测肾功能损伤指标；ELISA法和免疫组织化学法测定肾素-血管紧张素系统(renin-angiotensin system, RAS)组分表达；化学发光法测定肾组织超氧化物歧化酶(superoxide dismutase, SOD)、丙二醛(malondialdehyde, MDA)含量；免疫印迹法检测肾组织烟酰胺腺嘌呤二核苷酸磷酸(nicotinamide adenine dinucleotidephosphate, NADPH)氧化酶亚单位p47phox、诱导型一氧化氮合酶(inducible nitric oxide synthase, iNOS)、3-硝基酪氨酸(3-nitrotyrosine, 3-NT)、烟酰胺腺嘌呤二核苷酸磷酸氧化酶2 (NADPH oxidase 2, NOX2)、烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NADPH oxidase 4, NOX4)表达的变化。结果显示：与WT组相比，HT组小鼠血压明显升高；肾组织可见肾小球硬化，严重的间质、肾小管损伤，胶原沉积增多；24 h尿蛋白、血清肌酐和尿素水平上升；血清和肾组织中血管紧张素II (angiotensinII, Ang II)表达增加，血清血管紧张素-(1-7) [angiotensin-(1-7), Ang-(1-7)]表达降低，肾组织Ang-(1-7)表达增加；肾组织血管紧张素转化酶(angiotensin converting enzyme, ACE)、血管紧张素II 1型受体(angiotensin II type 1 receptor, AT1R)、Mas受体(Mas receptor, MasR)表达增加，ACE2表达减少；MDA含量增高，SOD含量下降，而且p47phox、iNOS、3-NT、NOX2、NOX4表达增加。阿利克仑可降低dTH小鼠血压，改善肾结构损伤和肾功能损伤，降低血清和肾组织Ang II及Ang-(1-7)水平，降低肾组织ACE和AT1R的表达，增加ACE2和MasR的表达，降低上述各项氧化应激指标水平。上述结果提示，阿利克仑可能通过调节ACE-Ang II-AT1R和ACE2-Ang-(1-7)-MasR轴之间的平衡、抑制氧化应激，对高血压肾损伤发挥保护作用。

关键词： 高血压; 阿利克仑; 肾素-血管紧张素系统; 肾素抑制剂; 肾损伤

Protective effect of aliskiren on renal injury in AGT-REN double transgenic hypertensive mice

YANG Xiao-Ling^1,2, CHEN Yan-Yan³, ZHAO Hua¹, ZHANG Bo-Yang¹, ZHANG Xiao-Fu¹, LI Xiao-Jie¹, YANG Xiu-Hong^1,2,*

¹School of Basic Medicine, North China University of Science and Technology, Tangshan 063000, China；²Hebei Key Laboratory of Basic Medicine for Chronic Diseases, Tangshan 063000, China；³Central Official Hospital, Beijing 100091, China

Abstract

This study aims to investigate the effects of renin inhibitor aliskiren on kidney injury in human angiotensinogen-renin (AGT-REN) double transgenic hypertensive (dTH) mice and explore its possible mechanism. The dTH mice were divided into hypertension group (HT group) and aliskiren intervention group (HT+Aliskiren group), while wild-type C57BL/6 mice were served as the control group (WT group). Blood pressure data of mice in HT+Aliskiren group were collected after 28 d of subcutaneous penetration of aliskiren (20 mg/kg), and the damage of renal tissue structure and collagen deposition were observed by HE, Masson and PAS staining. The ultrastructure of kidney was observed by transmission electron microscope. Coomassie bright blue staining and biochemical analyzer were used to detect renal function injury. The expression of renin-angiotensin system (RAS) was determined by ELISA and immunohistochemistry. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) in kidney were determined by chemiluminescence method. The content of nicotinamide adenine dinucleotide phosphate (NADPH) oxidase subunit p47phox, inducible nitric oxide synthase (iNOS), 3-nitrotyrosine (3-NT), NADPH oxidase 2 (NOX2) and NADPH oxidase 4 (NOX4) were detected by Western blot analysis. The results showed that compared with WT group, the blood pressure of mice in HT group was significantly increased. The renal tissue structure in HT group showed glomerular sclerosis, severe interstitial tubular injury, and increased collagen deposition. In addition, 24 h urinary protein, serum creatinine and urea levels increased. Serum and renal tissue levels of angiotensin II (Ang II) were increased, serum angiotensin-(1-7) [Ang-(1-7)] expression was decreased, and renal Ang-(1-7) expression was elevated. The expressions of ACE, Ang II type 1 receptor (AT1R) and MasR in renal tissue were increased, while the expression of ACE2 was decreased. MDA content increased, SOD content decreased, and the expressions of p47phox, iNOS, 3-NT, NOX2 and NOX4 were increased. However, aliskiren reduced blood pressure in dTH mice, improved renal structure and renal function, reduced Ang II and Ang-(1-7) levels in serum and renal tissue, reduced the expression of ACE and AT1R in renal tissue, increased the expression of ACE2 and MasR in renal tissue, and decreased the above levels of oxidative stress indexes in dTH mice. These results suggest that aliskiren may play a protective role in hypertensive renal injury by regulating the balance between ACEAng II-AT1R and ACE2-Ang-(1-7)-MasR axes and inhibiting oxidative stress.

Key words: Hypertension; aliskiren; renin-angiotensin system; renin inhibitor; kidney injury

收稿日期：　　录用日期：

通讯作者：杨秀红　　E-mail:

DOI: 10.13294/j.aps.2025.0046

引用本文：

杨晓玲, 陈妍妍, 赵华, 张博洋, 张晓伏, 李晓洁, 杨秀红. 阿利克仑对AGT-REN双转基因高血压小鼠肾损伤的保护作用[J]. 生理学报 2025; 77 (3): 408-418.

YANG Xiao-Ling, CHEN Yan-Yan, ZHAO Hua, ZHANG Bo-Yang, ZHANG Xiao-Fu, LI Xiao-Jie, YANG Xiu-Hong. Protective effect of aliskiren on renal injury in AGT-REN double transgenic hypertensive mice. Acta Physiol Sin 2025; 77 (3): 408-418 (in Chinese with English abstract).