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Research article
Rno-microRNA-30c-5p promotes myocardial ischemia reperfusion injury in rats through activating NF-κB pathway and targeting SIRT1
Shouyan Zhang
Luoyang Central Hospital Affiliated to Zhengzhou University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-4483-8131
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: This study aimed to investigate the regulatory effect of rno-microRNA-30c-5p (rno-miR-30c-5p) on myocardial ischemia reperfusion (IR) injury in rats and the underlying molecular mechanisms.
Methods: A rat model of myocardial IR injury was established. The infarct size was detected by 2,3,5-triphenyltetrazolium chloride staining. The pathologic changes of myocardial tissues were detected by hematoxylin-eosin staining. The apoptosis of myocardial cells was measured by TUNEL staining and flow cytometry. The mRNA expression of rno-miR-30c-5p and Sirtuin 1 (SIRT1) was detected by quantitative real-time PCR. The levels of IL-1β, IL-6 and TNF-α were detected by enzyme linked immunosorbent assay. The protein expression of Bax, Bcl-2, caspase-3, p-IκBα, IκBα, p-NF-κB p65, NF-κB p65 and SIRT1 was detected by Western blot. The interaction between rno-miR-30c-5p and SIRT1 was predicted by TargetScan, and further identified by dual luciferase reporter gene and RNA immunoprecipitation assay.
Results: The myocardial IR injury model was successfully established in rats. IR induced the myocardial injury in rats and increased the expression of rno-miR-30c-5p. Overexpression of rno-miR-30c-5p enhanced the inflammation, promoted the apoptosis, and activated NF-κB pathway in IR myocardial cells. SIRT1 was the target gene of rno-miR-30c-5p. Silencing of SIRT1 reversed the effects of rno-miR-30c-5p inhibitor on the apoptosis and NF-κB pathway in IR myocardial cells.
Conclusions: Rno-miR-30c-5p promoted the myocardial IR injury in rats through activating NF-κB pathway and down-regulating SIRT1.
Keywords
myocardial ischemia reperfusion injury, rno-miR-30c-5p, inflammation, apoptosis, SIRT1, NF-κB pathway
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On 24 Jan, 2020
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Editorial decision: Major revision
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Received 08 Nov, 2019
Reviewer #1 agreed
On 11 Oct, 2019
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Editor assigned
On 24 Sep, 2019
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Subject Areas
Cardiac & Cardiovascular Systems
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This preprint is under consideration at BMC Cardiovascular Disorders. A preprint is 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.
Learn more about In Review
Research article
Rno-microRNA-30c-5p promotes myocardial ischemia reperfusion injury in rats through activating NF-κB pathway and targeting SIRT1
Shouyan Zhang
Luoyang Central Hospital Affiliated to Zhengzhou University
Corresponding Author
ORCiD: https://orcid.org/0000-0003-4483-8131
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Under Review
Version 5
Posted 14 Apr, 2020
No community comments so far
Editor assigned
On 08 Apr, 2020
Submission checks complete
On 07 Apr, 2020
Editor invited
On 07 Apr, 2020
No comments provided
Editorial decision: Minor revision
On 01 Apr, 2020
Editor assigned
On 08 Mar, 2020
Submission checks complete
On 07 Mar, 2020
Editor invited
On 07 Mar, 2020
No comments provided
Editorial decision: Minor revision
On 03 Mar, 2020
Reviewer #1 agreed
On 02 Mar, 2020
Review #1 received
Received 02 Mar, 2020
Reviewers invited
Invitations sent on 01 Mar, 2020
Editor assigned
On 27 Feb, 2020
Submission checks complete
On 26 Feb, 2020
Editor invited
On 26 Feb, 2020
No comments provided
Editorial decision: Major revision
On 14 Feb, 2020
Review #2 received
Received 07 Feb, 2020
Review #1 received
Received 07 Feb, 2020
Reviewer #1 agreed
On 24 Jan, 2020
Reviewer #2 agreed
On 24 Jan, 2020
Reviewers invited
Invitations sent on 23 Jan, 2020
Editor assigned
On 22 Jan, 2020
Submission checks complete
On 21 Jan, 2020
Editor invited
On 21 Jan, 2020
No comments provided
Editorial decision: Major revision
On 22 Dec, 2019
Review #2 received
Received 02 Dec, 2019
Reviewer #2 agreed
On 18 Nov, 2019
Review #1 received
Received 08 Nov, 2019
Reviewer #1 agreed
On 11 Oct, 2019
Reviewers invited
Invitations sent on 09 Oct, 2019
Submission checks complete
On 24 Sep, 2019
Editor assigned
On 24 Sep, 2019
Editor invited
On 23 Sep, 2019
First submitted
On 20 Sep, 2019
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: This study aimed to investigate the regulatory effect of rno-microRNA-30c-5p (rno-miR-30c-5p) on myocardial ischemia reperfusion (IR) injury in rats and the underlying molecular mechanisms.
Methods: A rat model of myocardial IR injury was established. The infarct size was detected by 2,3,5-triphenyltetrazolium chloride staining. The pathologic changes of myocardial tissues were detected by hematoxylin-eosin staining. The apoptosis of myocardial cells was measured by TUNEL staining and flow cytometry. The mRNA expression of rno-miR-30c-5p and Sirtuin 1 (SIRT1) was detected by quantitative real-time PCR. The levels of IL-1β, IL-6 and TNF-α were detected by enzyme linked immunosorbent assay. The protein expression of Bax, Bcl-2, caspase-3, p-IκBα, IκBα, p-NF-κB p65, NF-κB p65 and SIRT1 was detected by Western blot. The interaction between rno-miR-30c-5p and SIRT1 was predicted by TargetScan, and further identified by dual luciferase reporter gene and RNA immunoprecipitation assay.
Results: The myocardial IR injury model was successfully established in rats. IR induced the myocardial injury in rats and increased the expression of rno-miR-30c-5p. Overexpression of rno-miR-30c-5p enhanced the inflammation, promoted the apoptosis, and activated NF-κB pathway in IR myocardial cells. SIRT1 was the target gene of rno-miR-30c-5p. Silencing of SIRT1 reversed the effects of rno-miR-30c-5p inhibitor on the apoptosis and NF-κB pathway in IR myocardial cells.
Conclusions: Rno-miR-30c-5p promoted the myocardial IR injury in rats through activating NF-κB pathway and down-regulating SIRT1.
Figure 1
Figure 2
Figure 3
Figure 4