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Long non-coding RNA KCNQ1OT1 increases the expression of PDCD4 by targeting miR-181a-5p, contributing to cardiomyocyte apoptosis in diabetic cardiomyopathy

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Abstract

Aims

Diabetic cardiomyopathy (DCM) is a specific myocardial alteration in patients with diabetics. LncRNA KCNQ1OT1 has been previously demonstrated to be involved in various diabetic complications. Our aims are to further investigate the underlying regulatory mechanisms/pathways of KCNQ1OT1 in DCM.

Methods

In vitro and in vivo models of DCM were established in high glucose (HG)-treated human cardiomyocytes and in streptozotocin (STZ)-induced diabetic mice, respectively. Gene and protein expressions were examined by qPCR, western blotting and ELISA. Cell proliferation and apoptosis were determined by CCK8 assay, flow cytometry and TUNEL staining. The association between KCNQ1OT1 and miR-181a-5p, miR-181a-5p and PDCD4 was predicted using bioinformatics methods and subsequently confirmed by dual luciferase reporter and RNA immunoprecipitation assays. Mouse cardiac tissues were collected and analysed using HE staining, Masson’s staining and immunohistochemical analysis.

Results

KCNQ1OT1 and PDCD4 were upregulated in HG-treated human cardiomyocytes, while miR-181a-5p was downregulated. In addition, KCNQ1OT1 could negatively regulate miR-181a-5p expression; meanwhile, miR-181a-5p also negatively regulated PDCD4 expression. KCNQ1OT1 silencing suppressed the expression of inflammatory cytokines and cell apoptosis in vitro, whereas inhibition of miR-181a-5p abrogated those effects of KCNQ1OT1 knockdown. Moreover, overexpressed PDCD4 abolished the inhibition on inflammation and apoptosis caused by miR-181a-5p overexpression. Finally, KCNQ1OT1 knockdown reduced the expression of PDCD4 via regulating miR-181a-5p and inhibited myocardial inflammation and cardiomyocyte apoptosis in the in vivo DCM model.

Conclusions

Our findings suggest that KCNQ1OT1 and its target gene miR-181a-5p regulate myocardial inflammation and cardiomyocyte apoptosis by modulating PDCD4 in DCM.

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Availability of data and materials

All data generated or analysed during this study are included in this article. The datasets used and/or analysed during the current study are available from the corresponding author on reasonable request.

Abbreviations

3′-UTR:

3′-Untranslated region

DCM:

Diabetic cardiomyopathy

HG:

High glucose

IL:

Interleukin

LncRNA:

Long non-coding RNA

miRNA:

MicroRNA

OD:

Optical density

qPCR:

Quantitative real-time PCR

sh:

Short-hairpin

STZ:

Streptozotocin

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments. The study was supported by Guangdong Natural Science Foundation (2018A030313535, Guangdong, China), Guangdong Medical Research Fund (A2020038) and Foundation for Basic and Applied Basic Research of Guangdong Province (2019A1515110063), Natural Science Foundation of Tibet Autonomous Region [XZ2020ZR-ZY55(Z)],National Key Research and Development Program (2018YFC2001803),National Natural Science Foundation of China(82002095).

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Author notes

  1. Equal contributors and co-first authors: Shuo-Fang Zhao and Ying-Xian Ye.

Authors and Affiliations

  1. Department of Anesthesiology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences and Guangdong Cardiovascular Institute, No.102, Zhongshan 2nd Road, Yuexiu District, Guangzhou, 510080, Guangdong Province, People’s Republic of China

    Shuo-Fang Zhao, Ying-Xian Ye, Jin-Dong Xu, Yi He, Deng-Wen Zhang & Sheng Wang

  2. Department of Anesthesiology, The University of Hong Kong, Pok Fu Lam, 999077, Hong Kong SAR, People’s Republic of China

    Zheng-Yuan Xia

  3. Department of Anesthesiology, Linzhi People’s Hospital, Linzhi, Tibet, People’s Republic of China

    Sheng Wang

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Contributions

SFZ helped in conception and study design; YXY and JDX contributed to data acquisition; YH and DWZ were involved in data analysis; ZYX helped in manuscript drafting; SW contributed to manuscript revising. All authors have read and approved the final version of this manuscript to be published.

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Correspondence to Sheng Wang.

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The authors declare that there is no conflict of interest.

Ethical approval

All animal study protocols approved by the Animal Ethics Committee of Guangdong Provincial People’s Hospital were performed in accordance with the guidelines by the National Institutes of Health Guide for the Care and Use of Laboratory Animals.

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Not applicable. This article does not contain any studies with human participants performed by any of the authors.

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The informed consent was obtained from the Animal Ethics Committee of Guangdong Provincial People’s Hospital.

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Zhao, SF., Ye, YX., Xu, JD. et al. Long non-coding RNA KCNQ1OT1 increases the expression of PDCD4 by targeting miR-181a-5p, contributing to cardiomyocyte apoptosis in diabetic cardiomyopathy. Acta Diabetol 58, 1251–1267 (2021). https://doi.org/10.1007/s00592-021-01713-x

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