miRNA-30e靶向5-LOX对大鼠脑缺血再灌注损伤的影响-东南大学学报医学版

miRNA-30e靶向5-LOX对大鼠脑缺血再灌注损伤的影响

单位：枝江市人民医院神经内科, 湖北枝江 443200

分类号：R-33

出版年·卷·期（页码）：2020·39·第六期（764-772）

摘要：

目的：探讨微小RNA-30e（miR-30e）通过5-脂氧合酶（5-LOX）对脑缺血再灌注（IR）损伤的作用。方法：将SD大鼠分为假手术组、模型组、载体组、miR-30e激动剂组、激动剂对照组、miR-30e拮抗剂组、拮抗剂对照组。2，3，5-氯化三苯基四氮唑染色法检测大鼠脑梗死体积；实时定量-聚合酶链反应检测组织中miR-30e的表达；蛋白质印迹实验检测5-LOX蛋白相对表达水平；酶联免疫吸附试验检测白三烯（CysLT）和白三烯B4（LTB4）表达水平。选取PC12细胞，分为常氧组、缺氧-葡萄糖剥夺和复氧模型（OGDR）组、mim-miR-NC组、mim-miR-30e组、mim-miR-30e+si-5-LOX组、inh-miR-NC组、inh-miR-30e组、inh-miR-30e+si-5-LOX组；MTT法检测细胞活力，流式细胞术检测细胞凋亡。结果：与假手术组比较，模型组大鼠神经功能评分、脑梗死体积、皮质梗死周围区组织miR-30e、5-LOX、CysLT和LTB4表达水平明显较高（P<0.05）；与激动剂对照组比较，miR-30e激动组上述指标明显较低（P<0.05）；与拮抗剂对照组比较，miR-30e拮抗剂组上述指标明显较高（P<0.05）；与常氧组比较，OGDR组miR-30e和5-LOX表达水平明显升高（P<0.05）；与mim-miR-NC组比较，mim-miR-30e组上述指标明显降低（P<0.05）；与inh-miR-NC组比较，inh-miR-30e组上述指标表达水平明显升高（P<0.05）；与常氧组比较，OGDR组细胞活力明显下降，而凋亡明显增加（P<0.05）；与mim-miR-NC组比较，mim-miR-30e组和mim-miR-30e+si-5-LOX组细胞活力明显升高，而凋亡明显减少（P<0.05）；与inh-miR-NC组比较，inh-miR-30e组细胞活力明显下降，而凋亡明显增加，inh-miR-30e+si-5-LOX组细胞活力明显升高，而凋亡明显减少（P<0.05）。结论：miR-30e通过抑制5-LOX表达对大鼠脑IR损伤具有潜在的神经保护作用。

Objective: To explore the effect of microRNA-30e(miR-30e) on cerebral ischemia-reperfusion(IR) injury through 5-lipoxygenase(5-LOX). Methods: SD rats were divided into the sham operation group, the model group, the vehicle group, the miR-30e agonist group, the agonist control group, the miR-30e antagonist group, and the antagonist control group. The 2,3,5-Chlorinated triphenyltetrazolium staining method was used to measure rat cerebral infarct volume; the real-time quantitative polymerase chain reaction method was used to measure the expression of miR-30e in tissues; Western blotting was used to measure the relative expression level of the 5-LOX protein; the enzyme-linked immunosorbent assay method was used to measure leukotriene(CysLT) and leukotriene B4(LTB4) expression levels. PC12 cells were divided into the normoxia group, the hypoxia-glucose deprivation and reoxygenation model(OGDR) group, the mim-miR-NC group, the mim-miR-30e group, the mim-miR-30e+si-5-LOX group, the inh-miR-NC group, the inh-miR-30e group, and the inh-miR-30e+si-5-LOX group. The MTT method was used to characterize cell viability; flow cytometry was used to characterize cell apoptosis. Results: Compared with those in the sham operation group, rats in the model group had significantly(P<0.05) higher neurological scores, cerebral infarction volumes, and expression levels of miR-30e, 5-LOX, CysLT and LTB4 in cortical infarction surrounding tissues(P<0.05). The above indicators of the miR-30e agonist group were significantly lower(P<0.05) compared with those of the agonist control group. The above indicators of the miR-30e antagonist group were significantly higher(P<0.05) compared with those of the antagonist control group. The expression levels of miR-30e and 5-LOX of the OGDR group were significantly increased(P<0.05) compared with those of the normoxia group. The above indicators of the mim-miR-30e group were significantly reduced(P<0.05) compared with those of the mim-miR-NC group. The expression levels of the above indicators of the inh-miR-30e group were significantly increased(P<0.05) compared with those of the inh-miR-NC group, the. The cell viability of the OGDR group was significantly decreased, while the apoptosis was significantly increased(P<0.05) compared with those of the normoxia group. The cell viability of mim-miR-30e group and mim-miR-30e+si-5-LOX group was significantly increased, while apoptosis was significantly decreased(P<0.05) compared with that of the mim-miR-NC group. The cell viability of the inh-miR-30e group significantly decreased, the apoptosis significantly increased, the cell viability of the inh-miR-30e+si-5-LOX group significantly increased, but the apoptosis significantly decreased(P<0.05) compared with the inh-miR-NC group. Conclusion: miR-30e has a potential neuroprotective effect on brain IR injury by inhibiting the expression of 5-LOX in rats.

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