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Paeonol suppresses the effect of ox-LDL on mice vascular endothelial cells by regulating miR-338-3p/TET2 axis in atherosclerosis

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Abstract

Atherosclerosis is the common vascular disease. Vascular smooth muscle cell proliferation and vascular endothelial cell (VEC) dysfunction are involved in the causes of atherosclerosis. And oxidized low-density lipoprotein (ox-LDL)-induced vascular endothelial cells (VECs) are suitable models for studying atherosclerosis development. Paeonol was reported to repress ox-LDL-induced VEC progression. However, its detailed mechanism was not fully reported. MicroRNAs (miRNAs) acted as regulators in multiple diseases. Previous findings found that microRNA-338-3p (miR-338-3p) was overexpressed in Atherosclerosis process. However, the function and underlying mechanism of miR-338-3p in ox-LDL-treated VECs needed to be elucidated. The purpose of this research was to reveal the role of miR-338-3p in paeonol-regulated ox-LDL-induced VEC progression. Cell counting kit-8 (CCK-8) and flow cytometry were employed to determine cell viability and apoptosis, respectively. Moreover, the levels of IL-6 and IL-1β were analyzed using enzyme-linked immunosorbent assay, as well as the contents of reactive oxygen species, lactate dehydrogenase, and malonic dialdehyde were investigated using related kits. Furthermore, quantitative real-time polymerase chain reaction was carried out to determine the expression of miR-338-3p. Western blot assay was conducted to detect the level of tet methylcytosine dioxygenase 2 (TET2). Besides, the interaction between miR-338-3p and TET2 was predicted by DIANA, and then confirmed by the dual-luciferase reporter assay and RNA immunoprecipitation assay. Ox-LDL repressed mice VEC viability, and promoted apoptosis, inflammatory response, and oxidative injury. Paeonol inhibited the effect of ox-LDL on the growth of the VECs. Furthermore, paeonol regulated VEC development via downregulating miR-338-3p expression. Interestingly, miR-338-3p targeted TET2 and inhibited TET2 expression. MiR-338-3p modulated ox-LDL-treated VEC growth through suppressing TET2 expression. We demonstrated that paeonol attenuated the effect of ox-LDL on the development of mice VECs via modulating miR-338-3p/TET2 axis, providing a theoretical basis for the treatment of AS.

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Abbreviations

AS:

Atherosclerosis

ox-LDL:

Oxidized low-density lipoprotein

VECs:

Vascular endothelial cells

TET2:

Tet methylcytosine dioxygenase 2

CCK-8:

Cell counting kit-8

PI:

Propidium iodide

V-FITC:

V-fluorescein isothiocyanate

ELISA:

Enzyme-linked immunosorbent assay

MDA:

Malonic dialdehyde

LDH:

Lactate dehydrogenase

ROS:

Reactive oxygen species

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

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  1. Department of Cardiology, Central China Fuwai Hospital, Heart Center of Henan Provincial People’s Hospital, No. 1 Yuwai Street, Zhengdong New District, Zhengzhou, 450000, Henan Province, China

    Yunfu Yu, Rui Yan, Xiaozhen Chen, Tao Sun & Jifeng Yan

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Yu, Y., Yan, R., Chen, X. et al. Paeonol suppresses the effect of ox-LDL on mice vascular endothelial cells by regulating miR-338-3p/TET2 axis in atherosclerosis. Mol Cell Biochem 475, 127–135 (2020). https://doi.org/10.1007/s11010-020-03865-w

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