Study on the Mechanism of LncRNA RMST Affects the Mechanism of Hydrogen Peroxide-Induced Cardiomyocyte Damage by Targeting miR-27a-3
XU Ling, XU Chaoxiang, LIN Xiaoxin, LU Huiyao, CAI Yinlian*
To explore the effect of LncRNA RMST on the damage of cardiomyocytes induced by hydrogen peroxide (H2O2) and its possible mechanism, H2O2 was used to induce rat cardiomyocyte H9C2 to establish a cell oxidative damage model. si-NC, si-LncRNA RMST, miR-NC, miR-27a-3p mimics, anti-miR-NC and si-LncRNA RMST, anti-miR-27a-3p and si-LncRNA RMST were transfected into H9C2 cells and added. The medium containing 200 μmol/L H2O2 was intervened for 2 h, which were recorded as si-NC+200 μmol/L H2O2 group, si-LncRNA RMST+200 μmol/L H2O2 group, miR-NC+200 μmol/L H2O2 group, miR-27a-3p+200 μmol/L H2O2 group, antimiR-NC+si-LncRNA RMST+200 μmol/L H2O2 group, anti-miR-27a-3p+si-LncRNA RMST+200 μmol/L H2O2 group. qRT-PCR method was used to detect the expression of LncRNA RMST and miR-27a-3p. The level of LDH in the cell culture fluid, as well as the levels of MDA and SOD in the cells were tested by using kits. Flow cytometry was used to detect the rate of apoptosis. The dual luciferase reporter experiment was used to detect the targeting relationship between LncRNA RMST and miR-27a-3p. Western blot method was used to detect the expression of Cleaved-caspase-3 and Bax protein. Compared with the si-NC+200 μmol/L H2O2 group, the levels of MDA and LDH in the si-LncRNA RMST+200 μmol/L H2O2 group were decreased (P<0.05), the rate of apoptosis and the protein levels of Cleaved-caspase-3 and Bax were decreased (P<0.05), while the level of SOD was increased (P<0.05). LncRNA RMST could target the expression of miR-27a-3p. Compared with the miR-NC+200 μmol/L H2O2 group, the levels of MDA and LDH in the miR-27a-3p+200 μmol/L H2O2 group were decreased (P<0.05), the rate of cell apoptosis and the protein levels of Cleaved-caspase-3, Bax were decreased (P<0.05), while the level of SOD was increased (P<0.05). Compared with the anti-miR-NC+si-LncRNA RMST+200 μmol/L H2O2 group, the levels of MDA and LDH in the anti-miR-27a-3p+si-LncRNA RMST+200 μmol/L H2O2 group were increased (P<0.05), the apoptosis rate and the protein levels of Cleaved-caspase-3 and Bax were increased (P<0.05), while the level of SOD was decreased (P<0.05). Interfering with the expression of LncRNA RMST could inhibit the oxidative stress response and apoptosis of cardiomyocytes by targeting miR-27a-3p, thereby reducing the damage of cardiomyocytes induced by H2O2.
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