miR-150-3p靶向CDH2基因对缺氧性肺动脉高压新生大鼠肺组织损伤的影响-现代医学

miR-150-3p靶向CDH2基因对缺氧性肺动脉高压新生大鼠肺组织损伤的影响

单位：海南省人民医院新生儿科, 海南海口 570100

分类号：R544.1;R563

出版年·卷·期（页码）：2022·50·第九期（1117-1123）

摘要：

目的：探究miR-150-3p靶向CDH2基因对缺氧性肺动脉高压（HPH）新生大鼠肺组织损伤的影响。方法：将新生Wistar大鼠随机分为对照组、模型组、阴性对照组和miR-150-3p过表达组，每组10只。除对照组外，其余各组构建新生大鼠HPH模型。分别检测各组大鼠缺氧3、6、12 d时的平均肺动脉压（mPAP），酶联免疫吸附测定法（ELISA）检测大鼠血清中缺氧诱导因子-1α（HIF-1α）和内皮素-1（ET-1）水平；在缺氧12 d时，Western blotting检测大鼠肺组织HIF-1α、ET-1、诱导型一氧化氮合酶（iNOS）和钙黏蛋白2（CDH2）表达情况，实时荧光定量PCR （RT-PCR）检测大鼠肺组织miR-150-3p、CDH2 mRNA表达。双荧光素酶报告基因实验验证miR-150-3p与CDH2之间的靶向关系。结果：与对照组相比，模型组和阴性对照组大鼠出现反应迟钝、萎靡不振、食量减少、肺组织炎症细胞浸润、肺泡大小改变等现象；大鼠mPAP，血清HIF-1α、ET-1水平，肺组织HIF-1α、ET-1、iNOS和CDH2蛋白表达均明显升高（P<0.05）；肺组织miR-150-3p表达明显降低，CDH2 mRNA表达明显升高（P<0.05）。与模型组、相比，阴性对照组上述指标均无显著差异（P>0.05）。与模型组、阴性对照组相比，miR-150-3p过表达组大鼠反应迟钝、炎症细胞浸润等不良现象均获得良好改善，且大鼠mPAP，血清HIF-1α、ET-1水平，肺组织HIF-1α、ET-1、iNOS和CDH2蛋白表达均降低（P<0.05）；肺组织miR-150-3p表达明显升高，CDH2 mRNA表达明显降低（P<0.05）。双荧光素酶报告基因实验结果证明CDH2为miR-150-3p的靶基因。结论：miR-150-3p能够通过靶向CDH2基因减轻HPH新生大鼠肺组织损伤。

Objective: To explore the effect of miR-150-3p targeting CDH2 gene on lung tissue damage in neonatal rats with hypoxia-induced pulmonary hypertension(HPH). Methods: Newborn Wistar rats were randomly divided into control group, model group, negative control group, and miR-150-3p overexpression group, with 10 rats in each group. Except for the control group, the rest of the groups constructed a newborn rat HPH model. The mean pulmonary arterial pressure(mPAP) of rats in each group was measured when hypoxia for 3, 6, and 12 d. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of hypoxia inducible factor-1α(HIF-1α) and endothelin-1(ET-1) in rat serum; on the 12th day of hypoxia, Western blotting was used to detect the expression of HIF-1α, ET-1, iNOS and CDH2 in the lung tissue of rats, RT-PCR was used to detect the expression of miR-150-3p and CDH2 mRNA in the lung tissue of rats. The dual luciferase reporter gene experiment was used to verify the targeting relationship between miR-150-3p and CDH2. Results: Compared with the control group, rats in the model group and the negative control group showed slow response, malaise, reduced appetite, lung tissue inflammatory cell infiltration, and alveolar size changes; mPAP, the levels of serum HIF-1α and ET-1, the expression of HIF-1α, ET-1, iNOS and CDH2 proteins in lung tissue were all significantly increased(P<0.05), the expression of miR-150-3p in lung tissue was significantly reduced, and the expression of CDH2 mRNA was significantly increased(P<0.05). Compared with the model group, there was no significant difference in the above indicators of the negative control group(P>0.05). Compared with the model group and the negative control group, the rats in the miR-150-3p overexpression group showed a good improvement in slow response and inflammatory cell infiltration; mPAP, the levels of serum HIF-1α and ET-1, the expression of HIF-1α, ET-1, iNOS and CDH2 proteins in lung tissue were all significantly decreased(P<0.05), the expression of miR-150-3p in lung tissue was significantly reduced, and the expression of CDH2 mRNA was significantly decreased(P<0.05). The results of the dual luciferase reporter gene experiment proved that CDH2 was the target gene of miR-150-3p. Conclusion: MiR-150-3p can reduce lung tissue damage in newborn rats with HPH by targeting CDH2 gene.

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