miR-30a-3p靶向NOD1对心肌细胞缺氧复氧损伤的影响

目的　探究miR-30a-3p与NOD1的靶向关系，及其对心肌细胞缺氧复氧损伤的影响和机制。 方法　将细胞分为Ctrl组、H/R组、miR-30a-3p mimic组和miR-30a-3p inhibitor组，经过缺氧复氧处理后，转染相应的miRNA处理细胞，RT-PCR检测miR-30a-3p和NOD1基因表达水平，荧光素酶报告检测miR-30a-3p和NOD1靶向关系，Western blot检测NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平，CCK8法检测细胞活性，Hoechst检测细胞凋亡，试剂盒检测LDH、CK、MDA、T-SOD水平。 结果　miR-30a-3p表达水平随缺氧复氧处理时间增长而下降，NOD1基因表达水平随缺氧复氧处理时间增长而升高。在荧光素酶报告实验中，NOD1 WT+miR-30a-3p mimic荧光素酶活性显著低于NOD1 WT+NC组。与Ctrl组比较，H/R组miR-30a-3p基因表达水平、细胞活性、T-SOD水平降低，NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率升高；与H/R组比较，miR-30a-3p mimic组miR-30a-3p基因表达水平、细胞活性、T-SOD水平升高，NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率降低，miR-30a-3p inhibitor组miR-30a-3p基因表达水平、细胞活性、T-SOD水平降低，NOD1、p-RPI2、p38 MAPK、NF-κB（p65）、cl-caspase-3蛋白表达水平、LDH、CK、MDA水平、细胞凋亡率升高。 结论　miR-30a-3p可靶向作用于NOD1，缓解心肌细胞缺氧复氧造成的损伤，其作用机制可能与调控NF-κB信号通路有关。

The effects of miR-30a-3p on Hypoxia/Reoxygenation induced injury of cardiomyocytes by targeting NOD1

Objective　To explore the targeting relationship between miR-30a-3p and NOD1, and its effect on Hypoxia/Reoxygenation induced injury of cardiomyocytes and its mechanism.　Methods　Cells were divided into Ctrl, H/R, miR-30a-3p mimic and miR-30a-3p inhibitor groups. After treatment with Hypoxia/Reoxygenation, cells in each group were transfected with corresponding miRNA. RT-PCR was performed for measuring the gene levels of miR-30a-3p and NOD1, luciferase reporter assay was performed for measuring the relationship between miR-30a-3p and NOD1, Western blot was used to determine the protein levels of NOD1, p-RPI2, p38 MAPK, NF-κB (p65), cl-caspase-3, cell viability was measured by CCK8, cell apoptosis was determined by Hoechst, and LDH, CK, MDA, T-SOD levels were detected by kit. Results　The expression level of miR-30a-3p decreased with the hypoxia-reoxygenation treatment time, and the expression level of NOD1 gene increased with the hypoxia-reoxygenation treatment time. In the luciferase reporter assay, the luciferase activity of NOD1 WT+miR-30a-3p mimic group was significantly lower than that of the NOD1 WTR+NC group. Compared with the Ctrl group, the gene level of miR-30a-3p, cell viability, T-SOD level were decreased, and the protein levels of NOD1, p-RPI2, p38 MAPK, NF-κB (p65), cl-caspase-3, expression levels of LDH, CK, MDA, cell apoptosis rate were increased in the H/R group. Compared with the H/R group, the gene level of miR-30a-3p, cell viability, T-SOD level were increased, and the protein levels of NOD1, p-RPI2, p38 MAPK, NF-κB（p65）, cl-caspase-3, expression levels of LDH, CK, MDA, and cell apoptosis rate were decreased in the miR-30a-3p mimic group. Meanwhile, the gene level of miR-30a-3p, cell viability, and T-SOD level were decreased, and the protein levels of NOD1, p-RPI2, p38 MAPK, NF-κB（p65）, cl-caspase-3, expression levels of LDH, CK, MDA, cell apoptosis rate were increased in the miR-30a-3p inhibitor group.　Conclusion　miR-30a-3p can target NOD1, alleviate the Hypoxia/Reoxygenation induced injury of carddiomyocytes, and the mechanism may be related to regulation of the NF-κB signaling pathway.