Protective Effect and Mechanism of miR-328-3p on Coronary Artery Endothelial Cell Injury Induced by Oxidized Low-density Lipoprotein

HOU Yonglan, LI Xia, WANG Jianmei, LIU Zhen, HAN Minglei, LIU Zhenghao, JIN Weidong

Abstract

To investigate the protective effect of miR-328-3p on oxidized low-density lipoprotein (ox-LDL)-induced coronary artery endothelial cell injury and the potentially relevant mechanisms.

Methods 

Human coronary artery endothelial cells (HCAECs) were induced with ox-LDL, and the cells were divided into a control group consisting of normal cells, an ox-LDL group receiving ox-LDL treatment, an ox-LDL+miR-NC group transfected with miR-NC and treated with ox-LDL, an ox-LDL+miR-328-3p group transfected with miR-328-3p and treated with ox-LDL, and ox-LDL+miR-328-3p+pcDNA group co-transfected miR-328-3p and pcDNA and treated with ox-LDL, and an ox-LDL+miR-328-3p+insulin-like growth factor 2 (IGF2) group co-transfected miR-328-3p and IGF2 and treated with ox-LDL. The expression level of miR-328-3p was determined with RT-qPCR. Cell proliferation was determined by MTT. Cell apoptosis was measured by flow cytometry. Western blot was conducted to examine the protein expression levels of cleaved cas-3 and IGF2. ELISA was performed to determine the levels of tumor necrosis factor α (TNF-α), interleukin (IL)-6, and IL-1β. Dual luciferase reporter experiment was performed to verify the targeting relationship between miR-328-3p and IGF2.

Results 

Compared with those of the control group, miR-328-3p expression level and cell activity were significantly reduced in the ox-LDL group (P<0.05), while the apoptotic rate, the protein expression levels of cleaved cas-3, IGF2, Bax, and Bcl-2, and the levels of TNF-α, IL-6, and IL-1β were significantly increased (P<0.05). Compared with those of the ox-LDL+miR-NC group, miR-328-3p expression level and cell activity significantly increased in the ox-LDL+miR-328-3p group (P<0.05), while the apoptosis rate, the protein expression levels of cleaved cas-3 and IGF2, and the levels of TNF-α, IL-6, and IL-1β were significantly reduced. IGF2 was a functional target of miR-328-3p. Compared with those of the ox-LDL+miR-328-3p+pcDNA co-transfection group, the IGF2 protein level was significantly increased (P<0.05) and cell activity was significantly decreased (P<0.05) in the ox-LDL+miR-328-3p+IGF2 co-transfection group, while the apoptosis rate, cleaved cas-3 protein level, and the levels of TNF-α, IL-6, and IL-1β were significantly elevated (P<0.05).

Conclusion 

miR-328-3p inhibits ox-LDL-induced apoptosis and inflammatory in coronary artery endothelial cell injury through targeted negative regulation of IGF2.

 

Keywords: miR-328-3p, Insulin-like growth factor 2, Oxidized low-density lipoprotein, Coronary artery endothelial cells

 

 

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References


FAN J, WATANABE T. Atherosclerosis: Known and unknown. Pathol Int, 2022, 72(3): 151–160. doi: 10.1111/pin.13202.

CAO H, JIA Q, YAN L, et al. Quercetin suppresses the progression of atherosclerosis by regulating MST1-mediated autophagy in ox-LDL-induced RAW264. 7 macrophage foam cells. Int J Mol Sci, 2019, 20(23): 6093. doi: 10.3390/ijms20236093.

CAI Y, XU L, XU C, et al. Hsa_circ_0001445 inhibits ox-LDL-induced HUVECs inflammation, oxidative stress and apoptosis by regulating miRNA-640. Perfusion, 2022, 37(1): 86–94. doi: 10.1177/0267 659120979472.

HE X N, XIN J Y, ZHAN J L, et al. Polycyclic aromatic hydrocarbons induce endothelial injury through miR-155 to promote atherosclerosis. Environ Mol Mutagen, 2021, 62(7): 409–421. doi: 10.1002/em.22454.

PAIM L R, SCHREIBER R, De ROSSI G, et al. Circulating microRNAs, vascular risk, and physical activity in spinal cord-injured subjects. J Neurotrauma, 2019, 36(6): 845–852. doi: 10.1089/neu.2018.5880.

ZHANG C, WANG L, SHEN Y. Circ_0004104 knockdown alleviates oxidized low-density lipoprotein-induced dysfunction in vascular endothelial cells through targeting miR-328-3p/TRIM14 axis in atherosclerosis. BMC Cardiovasc Disord, 2021, 21(1): 207. doi: 10.1186/s12872-021-02012-7.

XU H Y, ZHANG K L, WANG Y H, et al. The effect of Ox-LDL on the proliferation and expression of H19 and IGF2 in VSMCs. J Zunyi Med Univ, 2015, 38(4): 374–378.

MAO P, LIU X, WEN Y, et al. LncRNA SNHG12 regulates ox-LDL-induced endothelial cell injury by the miR-218-5p/IGF2 axis in atherosclerosis. Cell Cycle, 2021, 20(16): 1561–1577. doi: 10.1080/ 15384101.2021.1953755.

LIBBY P. The changing landscape of atherosclerosis. Nature, 2021, 592(7855): 524–533. doi: 10.1038/s41586-021-03392-8.

WATERBURY T M, TARANTINI G, VOGEL B, et al. Non-atherosclerotic causes of acute coronary syndromes. Nat Rev Cardiol, 2020, 17(4): 229–241. doi: 10.1038/s41569-019-0273-3.

KATTOOR A J, GOEL A, MEHTA J L. LOX-1: regulation, signaling and its role in atherosclerosis. Antioxidants (Basel), 2019, 8(7): 218. doi: 10. 3390/antiox8070218.

LI Z, ZHAO Y, SUGURO S, et al. MicroRNAs regulate function in atherosclerosis and clinical implications. Oxid Med Cell Longev, 2023, 2023: 2561509. doi: 10.1155/2023/2561509.

NOSALSKI R, SIEDLINSKI M, DENBY L, et al. T-cell-derived miRNA-214 mediates perivascular fibrosis in hypertension. Circ Res, 2020, 126(8): 988–1003. doi: 10.1161/CIRCRESAHA.119.315428.

YAO M, FANG C, WANG Z, et al. miR-328-3p targets TLR2 to ameliorate oxygen-glucose deprivation injury and neutrophil extracellular trap formation in HUVECs via inhibition of the NF-κB signaling pathway. PLoS One, 2024, 19(2): e0299382. doi: 10.1371/journal.pone. 0299382.

ZHAO H D, WU M L, CHEN P B, et al. Research progress of IGF2 gene expression regulation and its genetic variation in animal growth and development. Chin Husb Anim Vet Med, 2020, 47(6): 1844–1852.

KOU M S, LUO X L, GENG L Q. Expression of insulin-like growth factor-2 in coronary artery disease. Chin J Gen Pract, 2009, 7(1).

QU C, LIU X, HAN X, et al. miR-216b-5p regulates proliferation and apoptosis of ox-LDL-stimulated VSMCs and HUVECs via IGF2. J Biochem Mol Toxicol, 2023, 37(3): e23271. doi: 10.1002/jbt.23271.

YANG N, DONG B, SONG Y, et al. Downregulation of miR-637 promotes vascular smooth muscle cell proliferation and migration via regulation of insulin-like growth factor-2. Cell Mol Biol Lett, 2020, 25: 30. doi: 10.1186/s11658-020-00222-z.


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