Wang J, Chen X, Sun L, Chen X, Li H, Xiong B, Wang H
College of Basic Medical Sciences, Wannan Medical College, Wuhu 241002, China.
Graduate School, Wannan Medical College, Wuhu 241002, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Aug 20;42(8):1134-1142. doi: 10.12122/j.issn.1673-4254.2022.08.04.
To investigate the role of long non-coding RNA ZEB1-AS1 in cerebral ischemia/reperfusion injury (CI/RI).
We detected the temporal changes of ZEB1-AS1 and HMGB1 expression using qPCR and Western blotting in SD rats following CI/RI induced by middle cerebral artery occlusion (MCAO). The rat models of CI/RI were subjected to injections of vectors for ZEB1-AS1 overexpression or knockdown into the lateral ventricle, and the changes in cognitive function, brain water content, blood-brain barrier integrity, and IL-1β and TNF-α levels in the cerebrospinal fluid (CSF) and serum were observed. Neuronal loss and cell apoptosis in the cortex of the rat models were detected by FJC and TUNEL methods, and HMGB1 and TLR-4 expressions were analyzed with Western blotting. We also examined the effects of ZEB1-AS1 knockdown on apoptosis and expressions of HMGB1 and TLR-4 in SH-SY5Y cells with oxygen-glucose deprivation/reoxygenation (OGD/R).
In CI/RI rats, the expressions of ZEB1-AS1 and HMGB1 in the brain tissue increased progressively with the extension of reperfusion time, reaching the peak levels at 24 h followed by a gradual decline. ZEB1-AS1 overexpression significantly aggravated icognitive impairment and increased brain water content, albumin content in the CSF, and IL-1β and TNF-α levels in the CSF and serum in CI/RI rats ( < 0.05), while ZEB1-AS1 knockdown produced the opposite effects ( < 0.05 or 0.01). ZEB1-AS1 overexpression obviously increased the number of FJC-positive neurons in the cortex and enhanced the expressions of HMGB1 and TLR-4 in the rat models ( < 0.01); ZEB1-AS1 knockdown significantly reduced the number of FJC-positive neurons and lowered HMGB1 and TLR-4 expressions ( < 0.01). In SH-SY5Y cells with OGD/R, ZEB1-AS1 knockdown significantly suppressed cell apoptosis and lowered the expressions of HMGB1 and TLR-4 ( < 0.01).
ZEB1-AS1 overexpression aggravates CI/RI in rats through the HMGB1/TLR-4 signaling axis.
探讨长链非编码RNA ZEB1-AS1在脑缺血/再灌注损伤（CI/RI）中的作用。
我们采用qPCR和蛋白质免疫印迹法检测大脑中动脉闭塞（MCAO）诱导的CI/RI后SD大鼠中ZEB1-AS1和高迁移率族蛋白B1（HMGB1）表达的时间变化。对CI/RI大鼠模型侧脑室注射ZEB1-AS1过表达或敲低载体，观察认知功能、脑含水量、血脑屏障完整性以及脑脊液（CSF）和血清中白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）水平的变化。采用荧光金（FJC）和末端脱氧核苷酸转移酶介导的缺口末端标记（TUNEL）法检测大鼠模型皮质中的神经元丢失和细胞凋亡，并用蛋白质免疫印迹法分析HMGB1和Toll样受体4（TLR-4）的表达。我们还检测了ZEB1-AS1敲低对氧糖剥夺/复氧（OGD/R）处理的SH-SY5Y细胞凋亡以及HMGB1和TLR-4表达的影响。
在CI/RI大鼠中，脑组织中ZEB1-AS1和HMGB1的表达随再灌注时间延长而逐渐增加，在24小时达到峰值水平，随后逐渐下降。ZEB1-AS1过表达显著加重CI/RI大鼠的认知障碍，增加脑含水量、CSF中的白蛋白含量以及CSF和血清中的IL-1β和TNF-α水平（P<0.05），而ZEB1-AS1敲低则产生相反的效果（P<0.05或0.01）。ZEB1-AS1过表达明显增加大鼠模型皮质中FJC阳性神经元的数量，并增强HMGB1和TLR-4的表达（P<0.01）；ZEB1-AS1敲低显著减少FJC阳性神经元的数量，并降低HMGB1和TLR-4的表达（P<0.01）。在OGD/R处理的SH-SY5Y细胞中，ZEB1-AS1敲低显著抑制细胞凋亡，并降低HMGB1和TLR-4的表达（P<0.01）。
ZEB1-AS1过表达通过HMGB1/TLR-4信号轴加重大鼠的CI/RI。
