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Original investigation
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Background
Recent studies revealed that non-coding RNAs (ncRNAs) play a crucial role in pathophysiological processes involving diabetic cardiomyopathy that contributes to heart failure. The present study was designed to further investigate the anti-apoptotic effect of melatonin on cardiomyocyte in diabetic condition and to elucidate the potential mechanisms associated with ncRNAs.
Methods
In vivo, langendorff-perfusion system and histology staining were used to assess the effect of melatonin on cardiac function. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to measure the expression of ncRNAs. Protein expression levels were assessed by western bolt analysis. In vitro, hoechst 33258 staining and western bolt analysis were used to evaluate the effect of melatonin on apoptosis. We preformed luciferase assay and RNA immunoprecipitation to determine the targets of ncRNAs. RT-qPCR was used to observe the expression of ncRNAs in cardiomyocyte with high glucose treatment.
Results
In animal models, our results indicated that melatonin notably alleviated cardiac dysfunction and mitigated cardiomyocyte apoptosis in diabetic rats. Interestingly, lncRNA H19 level was increased along with concomitant decrease of miR-29c level in diabetic rats. However, we demonstrated that melatonin significantly downregulated H19 level and upregulated miR-29c level in vivo. In vitro experiments, it has been verified that positive modulation of miR-29c and inhibition of lncRNA H19 as well as mitogen-activated protein kinase (MAPK) pathways distinctly attenuated apoptosis in high glucose-treated H9c2 cells. Luciferase activity assay was conducted to evaluate the potential target sites of miR-29c on lncRNA H19 and MAPK13. LncRNA H19 silencing significantly downregulated the expression of the miR-29c target gene MAPK13 via inducing miR-29c expression. Furthermore, MAPK signal pathways were also affected through regulation of H19 and miR-29c. Most importantly, our results showed that melatonin alleviated hyperglycemic-induced cardiomyocyte apoptosis via inhibiting lncRNA H19/MAPK and increasing miR-29c level in vitro.
Conclusions
These results elucidate a novel protective mechanism of melatonin on diabetic cardiomyocyte apoptosis, which associated with the effect of melatonin on lncRNA H19/miR-29c expression and its downstream MAPK signal pathways, providing a promising strategy for preventing DCM in diabetic patients.
Keywords
melatonin, diabetic cardiomyopathy, apoptosis, non-coding RNA, MAPK
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A new preprint design is coming!
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Original investigation
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 10 May, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Cardiac & Cardiovascular Systems
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background
Recent studies revealed that non-coding RNAs (ncRNAs) play a crucial role in pathophysiological processes involving diabetic cardiomyopathy that contributes to heart failure. The present study was designed to further investigate the anti-apoptotic effect of melatonin on cardiomyocyte in diabetic condition and to elucidate the potential mechanisms associated with ncRNAs.
Methods
In vivo, langendorff-perfusion system and histology staining were used to assess the effect of melatonin on cardiac function. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was used to measure the expression of ncRNAs. Protein expression levels were assessed by western bolt analysis. In vitro, hoechst 33258 staining and western bolt analysis were used to evaluate the effect of melatonin on apoptosis. We preformed luciferase assay and RNA immunoprecipitation to determine the targets of ncRNAs. RT-qPCR was used to observe the expression of ncRNAs in cardiomyocyte with high glucose treatment.
Results
In animal models, our results indicated that melatonin notably alleviated cardiac dysfunction and mitigated cardiomyocyte apoptosis in diabetic rats. Interestingly, lncRNA H19 level was increased along with concomitant decrease of miR-29c level in diabetic rats. However, we demonstrated that melatonin significantly downregulated H19 level and upregulated miR-29c level in vivo. In vitro experiments, it has been verified that positive modulation of miR-29c and inhibition of lncRNA H19 as well as mitogen-activated protein kinase (MAPK) pathways distinctly attenuated apoptosis in high glucose-treated H9c2 cells. Luciferase activity assay was conducted to evaluate the potential target sites of miR-29c on lncRNA H19 and MAPK13. LncRNA H19 silencing significantly downregulated the expression of the miR-29c target gene MAPK13 via inducing miR-29c expression. Furthermore, MAPK signal pathways were also affected through regulation of H19 and miR-29c. Most importantly, our results showed that melatonin alleviated hyperglycemic-induced cardiomyocyte apoptosis via inhibiting lncRNA H19/MAPK and increasing miR-29c level in vitro.
Conclusions
These results elucidate a novel protective mechanism of melatonin on diabetic cardiomyocyte apoptosis, which associated with the effect of melatonin on lncRNA H19/miR-29c expression and its downstream MAPK signal pathways, providing a promising strategy for preventing DCM in diabetic patients.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8
Figure 9
Figure 10
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