俞艳华.长链非编码RNA NORA影响脂多糖诱导的心肌细胞炎症反应及细胞凋亡损伤的机制研究[J].内科急危重症杂志,2023,29(4):332-336
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| DOI：10.11768/nkjwzzzz20230417 |
| 中文关键词: 长链非编码RNA 微小RNA-378a-3p 脂多糖 心肌细胞 炎症反应 凋亡 |
| 英文关键词: |
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| 中文摘要: |
| 摘要 目的：探讨长链非编码RNA NORAD（LncRNA NORAD）是否通过调控微小RNA-378a-3p（miR-378a-3p）表达影响脂多糖（LPS）诱导的心肌细胞炎症反应及细胞凋亡。方法：体外培养大鼠心肌细胞H9C2，并将细胞分为6组：对照组、LPS组（10 μg/mL LPS）、LPS+阴性序列组（10 μg/mL LPS+转染阴性序列si-NC）、LPS+干扰NORAD组（10 μg/mL LPS+转染干扰NORAD）、LPS+干扰NORAD+ miR阴性对照组（10 g/mL LPS+共转染干扰NORAD和miR阴性对照）、LPS+干扰NORAD+ miR-378a-3p抑制剂组（10 μg/mL LPS+共转染干扰NORAD和miR-378a-3p抑制剂），除对照组细胞外，其余组细胞LPS刺激24 h。用实时荧光定量聚合酶链反应检测各组NORAD、miR-378a-3p相对表达量；用酶联免疫吸附试验（ELISA）检测白介素（IL）-6、肿瘤坏死因子（TNF）-α水平；以流式细胞术检测细胞凋亡率；以双荧光素酶报告实验验证NORAD与miR-378a-3p靶向结合关系。结果：与对照组比较，LPS组心肌细胞中NORAD的表达量、TNF-α、IL-6水平、细胞凋亡率升高，miR-378a-3p的表达量降低（P均＜0.05）；与LPS+阴性序列组比较，LPS+干扰NORAD组心肌细胞中TNF-α、IL-6水平降低及细胞凋亡率降低（P均＜0.05）；双荧光素酶报告实验证实NORAD可靶向结合miR-378a-3p；与LPS+干扰NORAD+ miR阴性对照组比较，LPS+干扰NORAD+miR-378a-3p抑制剂组心肌细胞中TNF-α、IL-6水平及细胞凋亡率升高（P均＜0.05）。结论：NORAD可通过调控miR-378a-3p减轻LPS诱导的心肌细胞炎症反应及细胞凋亡。 |
| 英文摘要: |
| Abstract Objective: To investigate whether LncRNA NORAD affects the inflammatory response and apoptosis of cardiomyocytes induced by lipopolysaccharide (LPS) through regulating the expression of microRNA-378a-3P (miR-378a-3p). Methods: Rat cardiomyocytes H9C2 were cultured in vitro, and si-NC, si-NORAD, si-NORAD and anti-miR-NC, si-NORAD and anti-miR-378a-3p were transfected into cardiomyocytes. Cardiomyocytes were stimulated with LPS for 24h. The relative expression of NORAD and miR-378a-3p was detected by real-time quantitative polymerase chain reaction (qRT-PCR). The levels of interleukin (IL)-6 and tumor necrosis factor (TNF)-α were detected by enzyme-linked immunosorbent assay (ELISA). Flow cytometry was used to detect the apoptosis rate. Dual-luciferase reporter assay was used to verify the targeting relationship between NORAD and miR-378a-3p. Results: Compared with the control group, the expression of NORAD in myocardial cells in the LPS group was significantly increased (P< 0.05), the expression of miR-378a-3p was significantly reduced (P< 0.05), and the levels of TNF-α and IL-6 were significantly increased (P< 0.05), apoptosis rate was increased significantly (P< 0.05). Compared with the LPS + si-NC group, the levels of TNF-α and IL-6 in cardiomyocytes of the LPS + si-NORAD group were significantly reduced (P <0.05), and the apoptosis rate was significantly reduced (P <0.05). Double luciferase reporting asssay confirmed that NORAD can target miR-378a-3p. Compared with the LPS + si-NORAD + anti-miR-NC group, the levels of TNF-α and IL-6 in cardiomyocytes of the LPS + si-NORAD + anti-miR-378a-3p group were significantly increased (P< 0.05), apoptosis rate was significantly increased (P< 0.05). Conclusion: NORAD attenuates LPS-induced cardiomyocyte inflammatory response and apoptosis by regulating miR-378a-3p. |
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