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Increased AT1 receptor expression mediates vasoconstriction leading to hypertension in Snx1−/− mice

Hypertension Research volume 44, pages 906–917 (2021)Cite this article

Abstract

Angiotensin II type 1 receptor (AT1R) is a vital therapeutic target for hypertension. Sorting nexin 1 (SNX1) participates in the sorting and trafficking of the renal dopamine D5 receptor, while angiotensin and dopamine are counterregulatory factors in the regulation of blood pressure. The effect of SNX1 on AT1R is not known. We hypothesized that SNX1, through arterial AT1R sorting and trafficking, is involved in blood pressure regulation. CRISPR/Cas9 system-generated SNX1−/− mice showed dramatic elevations in blood pressure compared to their wild-type littermates. The angiotensin II-mediated contractile reactivity of the mesenteric arteries and AT1R expression in the aortas were also increased. Moreover, immunofluorescence and immunoprecipitation analyses revealed that SNX1 and AT1R were colocalized and interacted in the aortas of wild-type mice. In vitro studies revealed that AT1R protein levels and downstream calcium signaling were upregulated in A10 cells treated with SNX1 siRNA. This may have resulted from decreased AT1R protein degradation since the AT1R mRNA levels showed no changes. AT1R protein was less degraded when SNX1 was downregulated, as reflected by a cycloheximide chase assay. Furthermore, proteasomal rather than lysosomal inhibition increased AT1R protein content, and this effect was accompanied by decayed binding of ubiquitin and AT1R after SNX1 knockdown. Confocal microscopy revealed that AT1R colocalized with PSMD6, a proteasomal marker, and the colocalization was reduced after SNX1 knockdown. These findings suggest that SNX1 sorts AT1R for proteasomal degradation and that SNX1 impairment increases arterial AT1R expression, leading to increased vasoconstriction and blood pressure.

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Acknowledgements

This study was supported in part by grants from the National Natural Science Foundation of China (31730043, 81570379), the National Key R&D Program of China (2018YFC1312700), the Program of Innovative Research Team by the National Natural Science Foundation (81721001), the Program for Changjiang Scholars and Innovative Research Team in University (IRT1216), The Third Affiliated Hospital of Chongqing Medical University (KY19024), and the National Institutes of Health (R01-DK039308, P01-HL074940).

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  1. These authors contributed equally: Chao Liu, Xingyue Li

Authors and Affiliations

  1. Department of Cardiology, Daping Hospital, The Third Military Medical University, Chongqing, China

    Chao Liu, Xingyue Li, Jinjuan Fu, Ken Chen, Qiao Liao, Jialiang Wang, Caiyu Chen, Hao Luo & Chunyu Zeng

  2. Chongqing Institute of Cardiology & Chongqing Key Laboratory of Hypertension Research, Chongqing, China

    Chao Liu, Jinjuan Fu, Ken Chen, Qiao Liao, Jialiang Wang, Caiyu Chen, Hao Luo & Chunyu Zeng

  3. Department of Emergency Medicine, Jinling Hospital, Medical School of Nanjing University, Nanjing, China

    Chao Liu

  4. College of Medicine, Southwest Jiaotong University, Chengdu, Sichuan, China

    Xingyue Li & Yongjian Yang

  5. Department of Cardiovascular Medicine, The General Hospital of Western Theater Command PLA, Chengdu, Sichuan, China

    Xingyue Li & Yongjian Yang

  6. Division of Renal Disease & Hypertension, The George Washington University School of Medicine and Health Sciences, Washington, DC, USA

    Pedro A. Jose

  7. Department of Clinical Nutrition, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, China

    Jian Yang

  8. State Key Laboratory of Trauma, Burns and Combined Injury, Daping Hospital, The Third Military Medical University, Chongqing, China

    Chunyu Zeng

  9. Cardiovascular Research Center of Chongqing College, Department of Cardiology of Chongqing General Hospital, University of Chinese Academy of Sciences, Chongqing, China

    Chunyu Zeng

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Liu, C., Li, X., Fu, J. et al. Increased AT1 receptor expression mediates vasoconstriction leading to hypertension in Snx1−/− mice. Hypertens Res 44, 906–917 (2021). https://doi.org/10.1038/s41440-021-00661-x

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