Downregulation of miR-3568 Protects Against Ischemia/Reperfusion-Induced Cardiac Dysfunction in Rats and Apoptosis in H9C2 Cardiomyocytes Through Targeting TRIM62

Abstract

microRNA-3568 (miR-3568) has been reported to be associated with atherosclerosis. Only few data describe the expression and underlying mechanism of miR-3568 in regulating cardiac ischemia-reperfusion (I/R) injury such as apoptosis. In this study, we therefore sought to investigate the potential function of miR-3568 in simulated I/R-induced apoptosis in H9C2 cardiomyocytes and related signaling pathways involved. Flow cytometry was performed to examine the cell apoptosis. The expression of miR-3568, Survivin, Bcl-2, ERK, JNK, p38, AKT, and STAT3 was measured by western blot and quantitative real-time PCR. The correlation between TRIM62 and p-STAT3 was measured by co-immunoprecipitation and ubiquitination. We found that miR-3568 expression in simulated I/R-induced H9C2 cardiomyocytes was increased in a time-dependent manner. miR-3568 mimic transfection in H9C2 cardiomyocytes significantly enhanced cell apoptosis, decreased the expression of Bcl-2 and Survivin, and activated STAT3 signaling, which were reversed by miR-3568 inhibitor. The direct interaction between miR-3568 and the 3′-untranslated region (UTR) of TRIM62 mRNA was confirmed by dualluciferase reporter assay. TRIM62 overexpression or AG490, a selective inhibitor of JAK2/ STAT3 significantly, significantly inhibited I/R and miR-3568 mimic induced cell apoptosis and STAT3 activation. TRIM62 was found to interact with and induce ubiquitination of p-STAT3. The facilitating role of miR-3568 in I/R injury was also observed in our in vivo rat models. In conclusion, our study suggests that miR-3568 promotes simulated I/Rinduced apoptosis in H9C2 cardiomyocytes through targeting TRIM62.

Identification of key microRNAs and the underlying molecular mechanism in spinal cord ischemia-reperfusion injury in rats

“…The expression of miR-3568 also increased in matrix vesicles (MV) compared with vascular smooth muscle cell (VSMC) in the rats with chronic kidney disease, indicating the role of miR-3568 in vascular calcification and/or MV formation ( Chaturvedi et al, 2015 ). A recent study revealed that miR-3568 expression in simulated IRI-induced H9C2 cardiomyocytes increased in a time-dependent manner, which promotes simulated IRI-induced apoptosis in H9C2 cardiomyocytes through targeting TRIM62 ( Li et al, 2020b ). Although miR-3568 was upregulated after SCII in rats based on microRNA microarrays results ( Chen et al, 2020b ; Li et al, 2015b ), the expression and potential function of miR-3568 in SCII has not been further explored.…”

“…Among the key DEmiRNAs not studied in SCII, miR-3568 was especially interesting. A recent study showed that miR-3568 markedly aggravated IRI-induced H9C2 cardiomyocytes apoptosis and decreased the expression of Bcl-2 and Survivin ( Li et al, 2020b ). This study implicated that miR-3568 might have antiapoptotic potentials.…”

“…A recent study revealed that miR-3568 expression in simulated IRI-induced H9C2 cardiomyocytes increased in a time-dependent manner, which promotes simulated IRI-induced apoptosis in H9C2 cardiomyocytes through targeting TRIM62 (Li et al, 2020b). Although miR-3568 was upregulated after SCII in rats based on microRNA microarrays results (Chen et al, 2020b;Li et al, 2015b), the expression and potential function of miR-3568 in SCII has not been further explored.…”

WTAP promotes myocardial ischemia/reperfusion injury by increasing endoplasmic reticulum stress via regulating m6A modification of ATF4 mRNA

“…Echocardiograms were recorded using a Vevo 770 high-resolution ultrasound imaging system (FUJIFILM VisualSonics, Toronto, ON, Canada) equipped with a dedicated micro-visualization scan-head probe (RMV-707B, single element probe) [ 33 ]. Rats were then euthanized and myocardium were collected for hematoxylin and eosin (HE) staining or TUNEL assay [ 34 ]. Animal Experimentation Ethics Committee of the Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine approved all experiments performed in this study.…”

Peer Review #1 of "Identification of key microRNAs and the underlying molecular mechanism in spinal cord ischemia-reperfusion injury in rats (v0.1)"

“…The expression of miR-3568 also increased in matrix vesicles (MV) compared with vascular smooth muscle cell (VSMC) in the rats with chronic kidney disease, indicating the role of miR-3568 in vascular calcification and/or MV formation (Chaturvedi et al 2015). A recent study revealed that miR-3568 expression in simulated IRI-induced H9C2 cardiomyocytes increased in a time-dependent manner, which promotes simulated IRI-induced apoptosis in H9C2 cardiomyocytes through targeting TRIM62 (Li et al 2020b). Although miR-3568 was upregulated after SCII in rats based on microRNA microarrays results (Chen et al 2020a;Li et al 2015b), the expression and potential function of miR-3568 in SCII has not been further explored.…”