【摘要】 目的 探究微小RNA-129-5p(miR-129-5p)靶向N-myc下游调控基因3(NDRG3)对脑出血(ICH)模型大鼠神经元损伤的影响。方法 大鼠脑内注射Ⅶ型胶原酶构建ICH模型，并将ICH模型大鼠随机分为ICH组、agomiR-NC组、agomiR-129-5p组、agomiR-129-5p+pCDH-NC组、agomiR-129-5p+pCDH-NDRG3组，每组均n=12；另取12只大鼠设为假手术组。mNSS评分评估各组大鼠神经功能缺损程度；苏木精-伊红染色观察各组脑组织病理变化；TUNEL染色检测各组神经元凋亡；qRT-PCR法检测各组大鼠脑组织中miR-129-5p表达水平；Western blot检测各组大鼠脑组织中NDRG3、Beclin-1、LC3蛋白表达；双荧光素酶报告基因实验检测miR-129-5p与NDRG3的靶向关系。结果 与假手术组比较，ICH组mNSS评分、神经元凋亡率、NDRG3 mRNA以及NDRG3、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达增高，miR-129-5p表达显著降低(均P<0.05);agomiR-129-5p组mNSS评分、神经元凋亡率、NDRG3mRNA以及NDRG3、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达较ICH组和agomiR-NC组降低，miR-129-5p表达较ICH组和agomiR-NC组增高(均P<0.05);agomiR-129-5p+pCDH-NDRG3组mNSS评分、神经元凋亡率、NDRG3 mRNA以及NDRG3、Beclin-1、LC3Ⅱ/Ⅰ蛋白表达较agomiR-129-5p组增高，miR-129-5p表达较agomiR-129-5p组和agomiR-129-5p+pCDH-NC组降低；双荧光素酶报告基因实验证实miR-129-5p可以靶向抑制NDRG3表达(P<0.05)。结论 miR-129-5p可能通过靶向抑制NDRG3表达改善ICH大鼠的脑神经元损伤。更多还原

【Abstract】 Aim To investigate the effect of microRNA-129-5p(miR-129-5p) on neuronal damage in intracerebral hemorrhage(ICH) rats by targeting N-myc downstream regulatory gene 3(NDRG3). Methods An ICH rat model was constructed by injecting type Ⅶ collagenase into the brain, and assigned into ICH group, agomiR-NC group, agomiR-129-5p group, agomiR-129-5p+pCDH-NC group, and agomiR-129-5p+pCDH-NDRG3 group randomly, with 12 rats in each group. Another 12 rats were labeled as sham surgery group. The mNSS score was performed to evaluate the degree of neurological deficits in each group. HE staining was used to observe the pathological changes in brain tissue of rats in each group. TUNEL staining was performed to detect neuronal apoptosis in each group. The qRT-PCR method was performed to detect miR-129-5p in the brain tissues in each group. Western blot was used to detect NDRG3, Beclin-1, and LC3 proteins in the brain tissues of rats in each group. Moreover, dual luciferase reporter gene assay was performed to detect the targeting relationship between miR-129-5p and NDRG3. Results The IH group had prominently higher mNSS score, apoptosis rate, NDRG3 mRNA, and NDRG3, Beclin-1, LC3 Ⅱ/Ⅰ protein expression, and prominently lower miR-129-5p expression than sham operation group(P<0.05). The agomiR-129-5p group had prominently lower mNSS score, apoptosis rate, NDRG3 mRNA, and NDRG3, Beclin-1, LC3 Ⅱ/Ⅰ protein expression, and prominently higher miR-129-5p expression than IH group and agomiR-NC group(P<0.05). The agomiR-129-5p+pCDH-NDRG3 group had prominently higher mNSS score, apoptosis rate, NDRG3 mRNA, and NDRG3, Beclin-1, LC3 Ⅱ/Ⅰ protein expression, and prominently lower miR-129-5p expression than agomiR-129-5p group and agomiR-129-5p+pCDH-NC group(P<0.05). And the dual luciferase reporter gene experiment confirmed that miR-129-5p could target and inhibit NDRG3 expression(P<0.05). Conclusion MiR-129-5p may improve neuronal damage in ICH rats by targeting the inhibition of NDRG3 expression.更多还原