Upregulation of miRNA-23a-3p rescues high glucose-induced cell apoptosis and proliferation inhibition in cardiomyocytes

Fang Wu,Yawen Zhang,Lian Liu,Xu Wang,Ke Cai,Jialing Zhang,Yiting Gui,Youhua Wang,Feng Wang,Yuanzheng Zheng,Yonghao Gui,Qiang Li,Qian Yang,Shuchun Li

doi:10.1007/s11626-020-00518-6

Abstract

Maternal hyperglycemia potentially inhibits the development of the fetal heart by suppressing cardiomyocyte proliferation and promoting apoptosis. Different studies have indicated that miRNAs are key regulators of cardiomyocyte proliferation, differentiation, and apoptosis and play a protective role in a variety of cardiovascular diseases. However, the biological function of miRNA-23a in hyperglycemia-related cardiomyocyte injury is not fully understood. The present study investigated the effect of miRNA-23a-3p on cell proliferation and apoptosis in a myocardial injury model induced by high glucose. H9c2 cardiomyocytes were exposed to high glucose to establish an in vitro myocardial injury model and then transfected with miRNA-23a-3p mimics. After miRNA-23a-3p transfection, lens-free microscopy was used to dynamically monitor cell numbers and confluence and calculate the cell cycle duration. CCK-8 and EdU incorporation assays were performed to detect cell proliferation. Flow cytometry was used to measured cell apoptosis. Upregulation of miRNA-23a-3p significantly alleviated high glucose-induced cell apoptosis and cell proliferation inhibition (p < 0.01 and p < 0.0001, respectively). The cell cycle of the miRNA-23a-3p mimics group was significantly shorter than that of the negative control group (p < 0.01). The expression of cell cycle–activating and apoptosis inhibition-associated factors Ccna2, Ccne1, and Bcl-2 was downregulated by high glucose and upregulated by miRNA-23a-3p overexpression in high glucose-injured H9c2 cells. miRNA-23a-3p mimics transfection before high glucose treatment had a significantly greater benefit than transfection after high glucose treatment (p < 0.0001), and the rescue effect of miRNA-23a-3p increased as the concentration increased. This study suggests that miRNA-23a-3p exerted a dose- and time-dependent protective effect on high glucose-induced H9c2 cardiomyocyte injury.

Highlights

Various embryo teratogenicity mechanisms are associated with maternal diabetes mellitus, such as heart defects and neural tube defects
Upregulation of miRNA-23a-3p rescued the high glucose (HG)-induced inhibition of cell proliferation The expression of miRNA-23a-3p in HG-treated H9c2 cells and normal glucose-treated cells was compared by qRT-PCR in this study, and the results showed that miRNA-23a-3p in HG-treated H9c2 cells was downregulated (p < 0.0001; Fig. 1a)
Since miRNA-23a-3p was downregulated in HG-treated H9c2 cells, we speculated that overexpression of miRNA-23a-3p may prevent HG-induced effects on H9c2 cells

Summary

Introduction

Various embryo teratogenicity mechanisms are associated with maternal diabetes mellitus, such as heart defects and neural tube defects. Maternal hyperglycemia might potentially inhibit cardiomyocyte proliferation and promote cell apoptosis during fetal heart development (Han et al 2015; Su et al 2016). Decreased cell proliferation and increased apoptosis are the two key factors leading to a reduction in the number of cardiomyocytes (Gutierrez et al 2009; Su et al 2017; Su et al 2019). Cardiomyocyte hypertrophy can compensate for the reduction in the number of cardiomyocytes, which is manifested as cardiac hypertrophy and subsequent heart failure (Ding et al 2013). Rescuing cardiomyocyte damage caused by hyperglycemia before birth is of great significance

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Conclusion