Title: Effect of miR-30b on cardiomyocyte apoptosis
Abstract: Objective
To investigate the effect of miR-30b on the apoptosis of rat H9C2 cardiomyocytes.
Methods
Rat H9C2 cells were divided into 3 groups: the blank control group, the negative control group and the miR-30b mimics group.The expression of miR-30b in 3 groups was detected by RT-PCR after LipofectamineTM2000 transfection.H9C2 cells were divided into 3 groups: the negative control group, the miR-30b mimics group and miR-30b mimics+ H2O2 group.The aApoptosis was detected 24 h after transfection, by Annexin V-FITC/PI double staining.and the Cleaved caspase-3, β-catenin, and c-myc protein were detected by Western blotting.
Results
The mRNA expression of miR-30b in the negative control group (1.007±0.062) was not statistically different from that in the blank control group (1.000±0.059)(F=0.200, P=0.849), but the miR-30b mRNA in the miR-30b mimics group (4.212±0.088) was significantly higher than that in the negative control group(F=72.929, P=0.000) and the blank control group(F=74.261, P=0.000). The apoptotic the negative control group, the miR-30b mimics group and the miR-30b mimics+ H2O2 group were (11.35±0.88)%, (1.22±0.13)%, and (5.83±0.41)%, respectively.The apoptoticsis rate cells in the miR-30b mimics and miR-30b mimics+ H2O2 groups was were significantly lower than that of the negative control group (F=778.088, P=0.000, F=193.976, P=0.000). The apoptotic cells rate in the of miR-30b mimics group was were significantly lower than that of miR-30b mimics + H2O2 group(F=689.257, P=0.000). The miR-30b mimics group and miR-30b mimics + H2O2 groups showed significantly lower levels of cleaved caspase-3(F=351.384, P=0.000, F=55.762, P=0.000), β-catenin expression(F=506.207, P=0.000, F=248.244, P=0.000), and c-myc protein(F=456.110, P=0.000, F=115.466, P=0.000) were significantly lower than that in the negative control group.The miR-30b mimics group showed significantly lower levels of Ccleaved caspase-3 (F=167.105, P=0.000), β-catenin(F=221.693, P=0.000), and c-myc(F=127.825, P=0.000)expression were significantly lower than that of the miR-30b mimics + H2O2 group(P<0.05).
Conclusion
Overexpression of miR-30b can decrease inhibit the apoptosis of H9C2 cardiomyocyte apoptosis, and in the mechanism is related to the down regulation of Wnt signaling pathway.
Key words:
Mir-30b; Cardiomyocytes; Apoptosis
Publication Year: 2018
Publication Date: 2018-07-05
Language: en
Type: article
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