Abstract：
The deregulation of long non-coding RNA (lncRNA) is associated with diverse human disorders, including cerebral ischemia/reperfusion injury (CI/RI). LncRNA SNHG14 was reported to function in CI/RI. Whereas, molecular mechanisms regulated by SNHG14 are not fully unveiled.
Mice subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) were used as CI/RI animal models. Neuro-2a (N2A) cells subjected to oxygen glucose deprivation/reoxygenation (OGD/R) were used as CI/RI cell models. The expression of SNHG14, miR-98-5p and BCL2 like 13 (BCL2L13) was examined using quantitative real-time PCR (qPCR) or western blot. Apoptosis was monitored by flow cytometry assay. Apoptosis-related markers and endoplasmic reticulum (ER) stress-related markers were quantified by western blot. Inflammatory factors and oxidative stress were detected using matched commercial kits. The predicted relationship between miR-98-5p and SNHG14 or BCL2L13 was validated by dual-luciferase reporter assay, RIP assay and pull-down assay.
The high expression of SNHG14 was monitored in MCAO/R-treated mice and OGD/R-treated N2A cells. OGD/R-induced N2A cell apoptosis, ER stress, inflammation and oxidative stress were attenuated by SNHG14 knockdown. SNHG14 targeted miR-98-5p to positively regulate BCL2L13 expression. Inhibition of miR-98-5p recovered cell apoptosis, ER stress, inflammation and oxidative stress that were repressed by SNHG14 knockdown. Overexpression of BCL2L13 enhanced cell apoptosis, ER stress, inflammation and oxidative stress that were repressed by miR-98-5p enrichment.
SNHG14 knockdown alleviated OGD/induced N2A cell apoptosis, ER stress, inflammation and oxidative stress by depleting BCL2L13 via increasing miR-98-5p.
Mice subjected to middle cerebral artery occlusion/reperfusion (MCAO/R) were used as CI/RI animal models. Neuro-2a (N2A) cells subjected to oxygen glucose deprivation/reoxygenation (OGD/R) were used as CI/RI cell models. The expression of SNHG14, miR-98-5p and BCL2 like 13 (BCL2L13) was examined using quantitative real-time PCR (qPCR) or western blot. Apoptosis was monitored by flow cytometry assay. Apoptosis-related markers and endoplasmic reticulum (ER) stress-related markers were quantified by western blot. Inflammatory factors and oxidative stress were detected using matched commercial kits. The predicted relationship between miR-98-5p and SNHG14 or BCL2L13 was validated by dual-luciferase reporter assay, RIP assay and pull-down assay.
The high expression of SNHG14 was monitored in MCAO/R-treated mice and OGD/R-treated N2A cells. OGD/R-induced N2A cell apoptosis, ER stress, inflammation and oxidative stress were attenuated by SNHG14 knockdown. SNHG14 targeted miR-98-5p to positively regulate BCL2L13 expression. Inhibition of miR-98-5p recovered cell apoptosis, ER stress, inflammation and oxidative stress that were repressed by SNHG14 knockdown. Overexpression of BCL2L13 enhanced cell apoptosis, ER stress, inflammation and oxidative stress that were repressed by miR-98-5p enrichment.
SNHG14 knockdown alleviated OGD/induced N2A cell apoptosis, ER stress, inflammation and oxidative stress by depleting BCL2L13 via increasing miR-98-5p.
摘要：
长链非编码RNA（lncRNA）的失调与多种人类疾病有关，包括脑缺血/再灌注损伤(CI/RI)。据报道，LncRNASNHG14在CI/RI中起作用。然而,SNHG14调控的分子机制尚未完全揭示。
将进行大脑中动脉闭塞/再灌注(MCAO/R)的小鼠用作CI/RI动物模型。将经受氧葡萄糖剥夺/复氧(OGD/R)的神经-2a(N2A)细胞用作CI/RI细胞模型。使用定量实时PCR(qPCR)或蛋白质印迹检查SNHG14、miR-98-5p和BCL2样13(BCL2L13)的表达。通过流式细胞术测定监测细胞凋亡。通过蛋白质印迹定量凋亡相关标志物和内质网(ER)应激相关标志物。使用匹配的商业试剂盒检测炎症因子和氧化应激。miR-98-5p与SNHG14或BCL2L13之间的预测关系通过双荧光素酶报告基因测定进行验证。RIP测定和下拉测定。
在MCAO/R处理的小鼠和OGD/R处理的N2A细胞中监测SNHG14的高表达。OGD/R诱导N2A细胞凋亡,ER压力,SNHG14敲低可减轻炎症和氧化应激。SNHG14靶向miR-98-5p以正向调节BCL2L13表达。抑制miR-98-5p恢复细胞凋亡，ER压力,SNHG14敲低抑制的炎症和氧化应激。过表达BCL2L13增强细胞凋亡,ER压力,miR-98-5p富集抑制的炎症和氧化应激。
SNHG14敲低减轻OGD/诱导的N2A细胞凋亡，ER压力,通过增加miR-98-5p消耗BCL2L13引起的炎症和氧化应激。
将进行大脑中动脉闭塞/再灌注(MCAO/R)的小鼠用作CI/RI动物模型。将经受氧葡萄糖剥夺/复氧(OGD/R)的神经-2a(N2A)细胞用作CI/RI细胞模型。使用定量实时PCR(qPCR)或蛋白质印迹检查SNHG14、miR-98-5p和BCL2样13(BCL2L13)的表达。通过流式细胞术测定监测细胞凋亡。通过蛋白质印迹定量凋亡相关标志物和内质网(ER)应激相关标志物。使用匹配的商业试剂盒检测炎症因子和氧化应激。miR-98-5p与SNHG14或BCL2L13之间的预测关系通过双荧光素酶报告基因测定进行验证。RIP测定和下拉测定。
在MCAO/R处理的小鼠和OGD/R处理的N2A细胞中监测SNHG14的高表达。OGD/R诱导N2A细胞凋亡,ER压力,SNHG14敲低可减轻炎症和氧化应激。SNHG14靶向miR-98-5p以正向调节BCL2L13表达。抑制miR-98-5p恢复细胞凋亡，ER压力,SNHG14敲低抑制的炎症和氧化应激。过表达BCL2L13增强细胞凋亡,ER压力,miR-98-5p富集抑制的炎症和氧化应激。
SNHG14敲低减轻OGD/诱导的N2A细胞凋亡，ER压力,通过增加miR-98-5p消耗BCL2L13引起的炎症和氧化应激。
