Department of Neurosurgery, Shanxian Central Hospital, Shanxian, 274300, Shandong, China.
Department of Nursing, Shanxian Central Hospital, No. 1 Wenhua Road, Shanxian, 274300, Shandong, China.
J Mol Neurosci. 2019 May;68(1):144-152. doi: 10.1007/s12031-019-01293-0. Epub 2019 Mar 20.
The objective of this study is to find out the potential influence of miR-301a in an experimental cerebral ischemia-reperfusion (I/R) rat model through targeting NDRG2. Rats with cerebral I/R injury were constructed and classified into model, miR-301a inhibitor, miR-301a mimic, NC (negative control), siNDRG2, NDRG2, and miR-301a inhibitor + si-NDRG2 groups, as well as another sham group. Cerebral infarct volume and cell apoptosis were observed by TTC staining and TUNEL staining. The targeting relationship between miR-301a and NDRG2 was verified by luciferase assay. ELISA, qRT-PCR, and Western blot were used to detect the expressions of related molecules. Compared with sham group, rats in the model group had elevated neurological function score and infarct volume; meanwhile, the cell apoptosis rate and inflammatory response were also increased with enhanced expression of miR-301a and NDRG2 (all P < 0.05). These changes were worsened in the miR-301a mimic and si-NDRG2 groups. Conversely, those rats in the miR-301a inhibitor and NDRG2 groups presented increased NDRG2, and at the same time, other above concerning factors also exhibited opposite tendencies (all P < 0.05). Dual-luciferase reporter gene assay confirmed that NDRG2 was a target gene of miR-301a, and si-NDRG2 could reverse the neuroprotective effect of miR-301a inhibitor in rats with cerebral I/R injury. Inhibiting miR-301a has a neuroprotective effect on rats with cerebral I/R injury to ameliorate cell apoptosis and inflammatory response through possibly targeting NDRG2.
本研究旨在通过靶向 NDRG2 研究 miR-301a 在实验性脑缺血再灌注（I/R）大鼠模型中的潜在影响。构建脑 I/R 损伤大鼠模型，并分为模型组、miR-301a 抑制剂组、miR-301a 模拟物组、NC（阴性对照组）组、siNDRG2 组、NDRG2 组、miR-301a 抑制剂+si-NDRG2 组以及另一个假手术组。通过 TTC 染色和 TUNEL 染色观察脑梗死体积和细胞凋亡情况。通过荧光素酶实验验证 miR-301a 与 NDRG2 的靶向关系。采用 ELISA、qRT-PCR 和 Western blot 检测相关分子的表达。与假手术组相比，模型组大鼠神经功能评分和梗死体积升高；同时，细胞凋亡率和炎症反应增加，miR-301a 和 NDRG2 的表达也增强（均 P<0.05）。miR-301a 模拟物和 siNDRG2 组的变化更为严重。相反，miR-301a 抑制剂和 NDRG2 组大鼠 NDRG2 表达增加，同时其他上述相关因素也呈现相反趋势（均 P<0.05）。双荧光素酶报告基因实验证实 NDRG2 是 miR-301a 的靶基因，si-NDRG2 可逆转 miR-301a 抑制剂对脑 I/R 损伤大鼠的神经保护作用。抑制 miR-301a 可能通过靶向 NDRG2 对脑 I/R 损伤大鼠发挥神经保护作用，改善细胞凋亡和炎症反应。
