Role of Lnc-MALAT1/miRNA-145/BNIP3 signaling pathway in sufentanil preconditioning-induced cardioprotection in rats: a cell experiment / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the role of long-chain non-coding RNA-lung cancer metastasis-related transcript 1/microRNA-145/Bcl-2 and adenovirus E1B19k Da interacting protein 3 (Lnc-MALAT1/miRNA-145/BNIP3) signaling pathway in sufentanil preconditioning-induced cardioprotection in rats.Methods:Rat H9C2 cells were inoculated in 6-well culture plates or flasks at a density of 1×10 6 cells/ml and divided into 5 groups ( n=30 each) using a random number table method: control group (group C), hypoxia-reoxygenation (H/R) group, sufentanil preconditioning group (S group), eukaryotic expression vector pcDNA3.0 group (pcDNA group) and pcDNA-MALAT1 group (MALAT1 group). Cells were incubated with 10 μmol/L sufentanil for 2 h, and then the H/R injury model was established in group S. In pcDNA group and MALAT1 group, cells were transfected with pcDNA3.0 and pcDNA-MALAT1, respectively, and then incubated with 10 μmol/L sufentanil for 2 h starting from 24 h after transfection, and then the H/R injury model was established.At 2 h after reoxygenation, the expression of Lnc-MALAT1, miRNA-145 and BNIP3 mRNA was detected by real-time polymerase chain reaction, the cell survival rate was detected by CCK-8, the apoptosis rate was detected by flow cytometry, the malondialdehyde (MDA) and superoxide dismutase (SOD) levels and amount of lactic dehydrogenase (LDH) released were detected, and the expression of Bcl-2, Bax and cleaved-caspase-3 was detected by Western blot. Results:Compared with group C, the survival rate was significantly decreased, apoptosis rate was increased, the MDA level and amount of LDH released were increased, SOD levels were decreased, the expression of LncRNA-MALAT1, BNIP3 mRNA, Bax, and cleaved-caspase-3 was up-regulated, and miRNA-145 and Bcl-2 expression was down-regulated in the other four groups ( P<0.05). Compared with group H/R, the cell survival rate was significantly increased, apoptosis rate was decreased, the MDA level and amount of LDH released were decreased, SOD levels were increased, the expression of LncRNA-MALAT1, BNIP3 mRNA, Bax, and cleaved-caspase-3 was down-regulated, and miRNA-145 and Bcl-2 expression was up-regulated in S and pcDNA groups ( P<0.05). Compared with group S, the survival rate was significantly decreased, apoptosis rate was increased, MDA level and amount of LDH released were increased, SOD levels were decreased, the expression of LncRNA-MALAT1, BNIP3 mRNA, Bax, and cleaved-caspase-3 was up-regulated, and miRNA-145 and Bcl-2 expression was down-regulated in group MALAT1 ( P<0.05). Conclusion:The mechanism of sufentanil preconditioning-induced cardioprotection is related to inhibiting Lnc-MALAT1/miRNA-145/BNIP3 signaling pathway in rats.