miR-133a-3p/TRPM4 axis improves palmitic acid induced vascular endothelial injury

Yadong Xue; Tingting Tong; Yuyao Zhang; Haijun Huang; Ling Zhao; Hongzhao Lv; Lingzhao Xiong; Kai Zhang; Yuxuan Han; Yuyang Fu; Yongzhen Wang; Rong Huo; Ning Wang; Tao Ban

doi:10.3389/fphar.2023.1340247

miR-133a-3p/TRPM4 axis improves palmitic acid induced vascular endothelial injury

Front Pharmacol. 2024 Jan 10:14:1340247. doi: 10.3389/fphar.2023.1340247. eCollection 2023.

Authors

Yadong Xue^#¹, Tingting Tong^#¹, Yuyao Zhang^#², Haijun Huang¹, Ling Zhao¹, Hongzhao Lv¹, Lingzhao Xiong¹, Kai Zhang¹, Yuxuan Han¹, Yuyang Fu¹, Yongzhen Wang¹, Rong Huo¹, Ning Wang¹, Tao Ban^{1

3

4

5}

Affiliations

¹ Department of Pharmacology (The Key Laboratory of Cardiovascular Research, Ministry of Education, State Key Laboratory of Frigid Zone Cardiovascular Diseases, Ministry of Science and Technology) at College of Pharmacy, Harbin Medical University, Harbin, China.
² Department of Anatomy, School of Basic Medical Sciences, Heilongjiang University of Chinese Medicine, Harbin, China.
³ Department of General Surgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, China.
⁴ Heilongjiang Academy of Medical Sciences, Harbin, China.
⁵ National-Local Joint Engineering Laboratory of Drug Research and Development of Cardio-Cerebrovascular Diseases in Frigid Zone, The National Development and Reform Commission, Harbin, China.

^# Contributed equally.

Abstract

Background: Vascular endothelial injury is a contributing factor to the development of atherosclerosis and the resulting cardiovascular diseases. One particular factor involved in endothelial cell apoptosis and atherosclerosis is palmitic acid (PA), which is a long-chain saturated fatty acid. In addition, transient receptor potential melastatin 4 (TRPM4), a non-selective cation channel, plays a significant role in endothelial dysfunction caused by various factors related to cardiovascular diseases. Despite this, the specific role and mechanisms of TRPM4 in atherosclerosis have not been fully understood. Methods: The protein and mRNA expressions of TRPM4, apoptosis - and inflammation-related factors were measured after PA treatment. The effect of TRPM4 knockout on the protein and mRNA expression of apoptosis and inflammation-related factors was detected. The changes of intracellular Ca²⁺, mitochondrial membrane potential, and reactive oxygen species were detected by Fluo-4 AM, JC-1, and DCFH-DA probes, respectively. To confirm the binding of miR-133a-3p to TRPM4, a dual luciferase reporter gene assay was conducted. Finally, the effects of miR-133a-3p and TRPM4 on intracellular Ca²⁺, mitochondrial membrane potential, and reactive oxygen species were examined. Results: Following PA treatment, the expression of TRPM4 increases, leading to calcium overload in endothelial cells. This calcium influx causes the assemblage of Bcl-2, resulting in the opening of mitochondrial calcium channels and mitochondrial damage, ultimately triggering apoptosis. Throughout this process, the mRNA and protein levels of IL-1β, ICAM-1, and VCAM1 significantly increase. Database screenings and luciferase assays have shown that miR-133a-3p preferentially binds to the 3'UTR region of TRPM4 mRNA, suppressing TRPM4 expression. During PA-induced endothelial injury, miR-133a-3p is significantly decreased, but overexpression of miR-133a-3p can attenuate the progression of endothelial injury. On the other hand, overexpression of TRPM4 counteracts the aforementioned changes. Conclusion: TRPM4 participates in vascular endothelial injury caused by PA. Therefore, targeting TRPM4 or miR-133a-3p may offer a novel pharmacological approach to preventing endothelial injury.

Keywords: MiR-133a-3p; TRPM4; hyperlipidemia; palmitic acid; vascular endothelial injury.

Grants and funding

The author(s) declare that no financial support was received for the research, authorship, and/or publication of this article. This work was supported by the National Natural Science Foundation of China (82070312, 82373868, 81872870), Scientific Fund of Heilongjiang Province (LH 2022H003), Scientific research project of Provincial Scientific Research Institute of Heilongjiang Province (CZKYF 2022-1-B007), Post-doctoral Research Start-up Fund Project of Heilongjiang Province (LBH-Q19155), Excellent Youth Fund of School of Pharmacy, Harbin Medical University (2019-YQ-01, 2020-YQ-02).