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6-Gingerol relieves myocardial ischaemia/reperfusion injury by regulating lncRNA H19/miR-143/ATG7 signaling axis-mediated autophagy

Abstract

Myocardial ischemia/reperfusion injury (MIRI) causes severe damage in cardiac tissue, thereby resulting in a high rate of mortality. 6-Gingerol (6-G) is reported to play an essential role in alleviating MIRI. However, the underlying mechanism remains obscure. This study was intended to explore the potential mechanism by which 6-G functions. Q-PCR was employed to quantify the relative RNA levels of long noncoding RNA (lncRNA) H19 (H19), miR-143, and ATG7, an enzyme essential for autophagy, in HL-1 cells. Western blotting, immunofluorescence, and immunohistochemistry were employed for protein evaluation in cultured cells or mouse tissues. Cell viability, cytotoxicity, and apoptosis were analysed by CCK-8, LDH, and flow cytometry assays, respectively. The binding sites for miR-143 were predicted using starBase software and experimentally validated through a dual-luciferase reporter system. Here, we found that 6-G elevated cellular H19 expression in hypoxia/reoxygenation (H/R)-treated HL-1 cells. Moreover, 6-G increased Bcl-2 expression but reduced cleaved caspase 3 and caspase 9 protein levels. Mechanistically, H19 directly interacted with miR-143 and lowered its cellular abundance by acting as a molecular sponge. Importantly, ATG7 was validated as a regulated gene of miR-143, and the depletion of miR-143 by H19 caused an increased in ATG7 expression, which in turn promoted the autophagy process. Last, mouse experiments highly supported our in vitro findings that 6-G relieves MIRI by enhancing autophagy. The H19/miR-143/ATG7 axis was shown to be critical for the function of 6-G in relieving MIRI.

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Fig. 1: 6-G attenuates H/R-induced cardiomyocyte injury and enhances autophagy.
Fig. 2: 6-G attenuates myocardial injury by relying on enhanced autophagy.
Fig. 3: 6-G promotes autophagy by upregulating H19 and relieves cardiomyocyte injury caused by H/R.
Fig. 4: H19 acts as a ceRNA by absorbing miR-143 and indirectly upregulates ATG7 expression.
Fig. 5: The H19/miR-143/ATG7 axis is required for 6-G to alleviate H/R injury.
Fig. 6: 6-G relieves myocardial ischemia-reperfusion injury through promotion of autophagy in mice.

Data availability

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

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Acknowledgements

We would like to give our sincere gratitude to the reviewers for their constructive comments.

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Conception and study design: Xiang-Wei Lv; Data acquisition: Meng-Jie Wang; Data analysis: Qiu-Yu Qin; Manuscript drafting: Pan Lu; Manuscript revising: Guo-Wei Qin.

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Correspondence to Guo-Wei Qin.

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The animal ethics involved in the experiment was approved by the Guilin Medical University Laboratory Animal Ethics Committee.

Funding

This study was supported by grants from the Natural Science Foundation of Guangxi (no. 2018GXNSFDA281039) and Natural Science Foundation of Guangxi (no.2018GXNSFAA294096).

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Lv, XW., Wang, MJ., Qin, QY. et al. 6-Gingerol relieves myocardial ischaemia/reperfusion injury by regulating lncRNA H19/miR-143/ATG7 signaling axis-mediated autophagy. Lab Invest (2021). https://doi.org/10.1038/s41374-021-00575-9

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