• 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
  • 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
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    俞艳华 枣庄矿业集团枣庄医院 c8zb8z@163.com 
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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.