Cerebral ischemia–reperfusion injury (CI/RI) is one of the most common diseases of the central nervous system. At present, there is no specific treatment for CI/RI. It is necessary to explore the mechanism of CI/RI and find new ways to prevent and treat CI/RI. An oxygen and glucose deprivation/recovery (OGD/R) model was established to evaluate the effects of mouse astrocytes (MA-C) cell viability and apoptosis of stepwise exposure to oxygen and glucose deprivation followed by their replenishment. This assessment included using taurine upregulated gene 1–small interfering RNAs (TUG1-siRNA) transfection to determine the effects of TUG1 knockdown on MA-C survival and apoptosis. Real-time quantitative polymerase chain reaction (RT-qPCR) was used to evaluate TUG1 and miR-142-3p expression levels. The luciferase gene reporter assay was performed to validate that miR-142-3p is a TUG1 target. Accordingly, the effects of miR-142-3p knockdown on TUG1-induced MA-C apoptosis were determined using flow cytometry. Methyl thiazolyl tetrazolium (MTT) method was used to detect cell growth viability. Western blotting analysis was performed to detect the expression levels of apoptosis-related proteins. TUG1 was upregulated, while miR-142-3p was downregulated in the OGD/R model of MA-C cells. Inhibiting the expression of TUG1 could protect MA-C cells and reverse the decrease in growth viability and increasing apoptosis of MA-C cells caused by OGD/R stimulation. On the other hand, the inhibition of miR-142-3p offset the effect of TUG1 knockdown on cell viability and apoptosis. Inhibition of OGD/R-induced increases in TUG1 expression that in turn reduces miR-142-3p upregulation may suppress reperfusion-induced losses in cell viability.

中文翻译：

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敲除lncRNA TUG1通过调节miR-142-3p减轻脑缺血/再灌注损伤

脑缺血再灌注损伤（CI/RI）是中枢神经系统最常见的疾病之一。目前尚无针对CI/RI的特异性治疗方法。有必要探索CI/RI的发病机制，寻找防治CI/RI的新途径。建立了氧气和葡萄糖剥夺/恢复 (OGD/R) 模型，以评估小鼠星形胶质细胞 (MA-C) 细胞活力和逐步暴露于氧气和葡萄糖剥夺然后补充它们的细胞凋亡的影响。该评估包括使用牛磺酸上调基因 1-小干扰 RNA (TUG1-siRNA) 转染来确定 TUG1 敲低对 MA-C 存活和细胞凋亡的影响。实时定量聚合酶链反应 (RT-qPCR) 用于评估 TUG1 和 miR-142-3p 表达水平。进行荧光素酶基因报告分析以验证 miR-142-3p 是 TUG1 靶标。因此，使用流式细胞术确定 miR-142-3p 敲低对 TUG1 诱导的 MA-C 细胞凋亡的影响。甲基噻唑基四唑（MTT）法用于检测细胞生长活力。进行蛋白质印迹分析以检测凋亡相关蛋白的表达水平。在 MA-C 细胞的 OGD/R 模型中，TUG1 上调，而 miR-142-3p 下调。抑制 TUG1 的表达可以保护 MA-C 细胞并逆转由 OGD/R 刺激引起的 MA-C 细胞生长活力的降低和细胞凋亡的增加。另一方面，miR-142-3p 的抑制抵消了 TUG1 敲低对细胞活力和细胞凋亡的影响。