Small ubiquitin-like modifiers inhibitors lower blood pressure via ERK5/KLF2-dependent upregulation of the eNOS/NO pathway

Abstract

Background Small ubiquitin-like modifiers (SUMO)ylation is a dynamic and reversible post-translational modification playing pivotal roles in the regulation of cancer, diabetes, heart failure, and neurological diseases. However, whether SUMO inhibitors also have anti-hypertension effect remains yet to be explored. Methods Blood pressure was monitored in spontaneously hypertensive rats (SHR) after Tannic acid (TA) administration for 4 weeks. The contents of nitric oxide (NO) and endothelin-1 (ET-1) in the serum of SHR were measured. Isolated endothelium-intact mesenteric artery rings were used to study relaxation effect of SUMO inhibitors. ERK5 SUMOylation was determined using coimmunoprecipitation (co-IP) and immunofluorescence (IF). NO levels were analyzed by IF. The expression levels of KLF2 and p-eNOS were semiquantified by Western blot analysis. The transcriptional activity of eNOS promotor was assayed using ChIP-PCR. Results Three SUMO inhibitors all reduced the phenylephrine (PE)-induced contraction of mesenteric artery rings in a concentration-dependent manner. Co-IP revealed that ponatinib promoted ERK5 SUMOylation, which was nulled following pretreatment with the SUMO inhibitors. IF displayed that TA increased ERK5 accumulation and its co-localization with SUMO-1 in the nucleus. ChIP-PCR unveiled TA-induced enhancement of KLF2-dependent eNOS promoter activity and upregulation of eNOS/NO expression in HUVECs. In vivo, TA significantly lowered the blood pressure and improved the vascular reactivity by activating the KLF2/eNOS/NO pathway. Additionally, the level of NO was elevated along with decreased ET-1 levels in the serum of SHR. Conclusion SUMO inhibitors inhibit ERK5 SUMOylation to promote KLF2-eNOS/NO signaling, indicating their therapeutic potential for the treatment of hypertension.

Keywords

hypertension / SUMO inhibitor / ERK5 / SUMOylation / KLF2

Author contributions

Yang B F, Zhao D and Liu Y designed the experiments and supervised the project. Tang N N, Li J T and Wang Z were responsible for the manuscript writing. Zuo J L and Zhang Z F analyzed the data. Sun Y L, Li X, Mu R X, Huang D, Han Y N and Chen Y Q performed the in vitro experiments. Ma Q X, Zhang J, Wu J Y, Wang H, Zhao H X and Dong X L conducted the in vivo experiments. Wang Z G revised the manuscript.

Source of funding

This study was supported by the National Natural Science Foundation of China (82330011, U21A20339), the CAMS Innovation Fund for Medical Sciences (CIFMS, 2020-I2M-5-003), the National Natural Science Foundation of China (No. 82370302, 31871175), Natural Science Foundation of Heilongjiang Province of China (No. YQ2019H003), College of Pharmacy, Harbin Medical University Excellent Young Talents Funding (No. 2020-YQ-01).

Ethics approval

All animal experiments were conducted in accordance with the guidelines of the American Association for the Accreditation of Laboratory Animal Care International and the Guide for the Care and Use of Laboratory Animals published by the US National Institutes of Health (NIH Publication No. 85-23, revised 1996). All procedures were approved by the Institutional Animal Care and Use Committee of Harbin Medical University (Ethical approval number: IRB3017723).

Conflict of interest

Yang B F is the editor-in-chief of Frigid Zone Medicine. The article was subject to the journal's standard procedures, with peer review handled independently of him and his research groups.

Data availability statement

All data in this studyare available from from the corresponding author, on request.

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