马钱子苷通过LncRNA MALAT1靶向microRNA-155-5p促进雪旺细胞增殖和迁移的机制研究

Loganin promotes Schwann cell proliferation and migration by targeting microRNA-155-5p through lncRNA MALAT1

摘要:

目的 探究马钱子苷（Loganin）通过LncRNA MALAT1靶向microRNA-155-5p（miR-155-5p）促进雪旺细胞增殖和迁移的机制研究。方法 以小鼠雪旺细胞为研究对象，实验设置对照组（oe-NC组、si-NC组、Control组）、LncRNA MALAT1过表达组（oe-MALAT1组）、LncRNA MALAT1干扰组（si-MALAT组）、共转染oe-MALAT1 + mimic NC组或oe-MALAT1 + miR mimic（miR-155-5p过表达）组；用50、100和200 μmol/L浓度的Loganin处理细胞，分为Loganin（50 μmol/L）组、Loganin（100 μmol/L）组、Loganin（200 μmol/L）组；细胞转染si-NC或si-MALAT后用100 μmol/L Loganin处理，分为Loganin（100 μmol/L）+ si-NC组和Loganin（100 μmol/L）+ si-MALAT组。通过实时荧光聚合酶链反应（qRT-PCR）检测雪旺细胞中LncRNA MALAT1和miR-155-5p的表达；利用CCK-8实验和Transwell实验检测oe-NC组、oe-MALAT1组、oe-MALAT1+ mimic NC组及oe-MALAT1 + miR mimic组雪旺细胞增殖和迁移能力。通过荧光素酶报告实验验证LncRNA MALAT1与miR-155-5p的靶向关系。qRT-PCR检测不同浓度的Loganin（50、100和200 μmol/L）对雪旺细胞中LncRNA MALAT1表达的影响。通过CCK-8实验和Transwell实验检测不同浓度的Loganin组、Loganin（100 μmol/L）+ si-NC组和Loganin（100 μmol/L）+ si-MALAT组中雪旺细胞增殖和迁移的能力。结果 与oe-NC组比较，oe-MALAT1组中LncRNA MALAT1水平升高（P <0.05）；且oe-MALAT1组雪旺细胞的增殖和迁移能力增强（P <0.05）。荧光素酶报告实验证实LncRNA MALAT1靶向负调节miR-155-5p，抑制MALAT1的水平可促进miR-155-5p的表达（P <0.05）。与oe-MALAT1 + mimic NC组组比较，oe-MALAT1 + miR mimic组雪旺细胞增殖和迁移能力减弱（P <0.05）。与Control组比较，不同浓度的Loganin促进雪旺细胞增殖和迁移（P <0.05），且具有浓度效应。此外，Loganin显著促进LncRNA MALAT1的表达（P <0.05）。相对于Loganin（100 μmol/L）+ si-NC组，Loganin（100 μmol/L）+ si-MALAT组雪旺细胞的增殖和迁移能力被抑制（P <0.05）。结论 Loganin通过上调LncRNA MALAT1而抑制miR-155-5p的表达，促进雪旺细胞的增殖和迁移，为坐骨神经损伤的治疗提供了新的方向。

Abstract:

Objective To explore the whether loganin promotes Schwann cell proliferation and migration by targeting microRNA-155-5p through lncRNA MALAT1.Methods Using mouse Schwann cells as the research model, the experiment was designed with the following groups: control groups (oe-NC and si-NC groups), lncRNA MALAT1 overexpression group (oe-MALAT1 group), lncRNA MALAT1 interference group (si-MALAT1 group), oe-MALAT1 + mimic NC co-transfection group (oe-MALAT1 + mimic NC group) and oe-MALAT1 + miR mimic co-transfection group (miR-155-5p overexpression group). Cells were treated with different concentrations of loganin (50, 100, and 200 μmol/L) and divided into loganin (50 μmol/L), loganin (100 μmol/L), and loganin (200 μmol/L) groups, and cells transfected with si-NC or si-MALAT were treated with 100 μmol/L loganin and divided into loganin (100 μmol/L) + si-NC group and loganin (100 μmol/L) + si-MALAT group. The expression levels of lncRNA MALAT1 and miR-155-5p in Schwann cells were detected by quantitative real-time polymerase chain reaction (qRT-PCR), and the proliferation and migration abilities of Schwann cells in the oe-NC group, the oe-MALAT1 group, the oe-MALAT1 + mimic NC group, and the oe-MALAT1 + miR mimic group were determined using the CCK-8 assay and the Transwell assay. The targeting relationship between lncRNA MALAT1 and miR-155-5p was verified by luciferase reporter assay. The qRT-PCR was performed to detect the effects of different concentrations of loganin (50, 100, and 200 μmol/L) on the expression of lncRNA MALAT1 in Schwann cells. The ability of Schwann cells to proliferate and migrate in different concentrations of loganin, and that in the loganin (100 μmol/L) + si-NC group and the loganin (100 μmol/L) + si-MALAT group, were detected using the CCK-8 assay and the Transwell assay.Results Compared with the oe-NC group, the level of lncRNA MALAT1 in the oe-MALAT1 group increased significantly (P < 0.05). Moreover, the proliferation and migration abilities of Schwann cells in the oe-MALAT1 group were significantly enhanced (P < 0.05). The luciferase reporter assay confirmed that lncRNA MALAT1 targeted and negatively regulated miR-155-5p, and inhibiting the level of MALAT1 significantly promoted the expression of miR-155-5p (P < 0.05). Compared with the oe-MALAT1 + mimic NC group, the proliferation and migration abilities of Schwann cells in the oe-MALAT1 + miR mimic group were weakened (P < 0.05). Compared with the control group, different concentrations of loganin significantly promoted the proliferation and migration of Schwann cells (P < 0.05), exhibiting a dose-dependent effect. In addition, loganin significantly promoted the expression of lncRNA MALAT1 (P < 0.05). Compared with the loganin (100 μmol/L) + si-NC group, the proliferation and migration of Schwann cells in the loganin (100 μmol/L)+ si-MALAT group were significantly inhibited (P < 0.05).Conclusions By up-regulating lncRNA MALAT1, loganin inhibits the expression of miR-155-5p and promotes the proliferation and migration of Schwann cells, providing a novel insight for the treatment of sciatic nerve injury.