Department of Neurosurgery & Brain and Nerve Research Laboratory, First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.
Institute of Stroke Research, Soochow University, Suzhou, Jiangsu, China.
CNS Neurosci Ther. 2022 Jul;28(7):1124-1138. doi: 10.1111/cns.13850. Epub 2022 Apr 25.
The aim was to study the role of Rph3A in neuronal injury induced by cerebral ischemia-reperfusion.
The protein and mRNA levels of Rph3A in penumbra were detected by Western blot. The localization of Rph3A in different cell types in penumbra was detected by immunofluorescence. Apoptosis in the brain was detected by TUNEL staining. We tested neurobehavioral evaluation using rotarod test, adhesive-removal test, and Morris Water maze test. We examined the expression and localization of Rph3A in cultured neurons and astrocytes in vitro by Western blot and ELISA, respectively.
The mRNA and protein levels of Rph3A had significantly increased in brain penumbra of the rat MCAO/R model. Rph3A was mainly distributed in neurons and astrocytes and was significantly increased by MCAO/R. We downregulated Rph3A and found that it further worsened the cerebral infarct, neuronal death and behavioral, cognitive, and memory impairments in rats after MCAO/R. We also found that ischemia-reperfusion upregulated the in vitro protein level and secretion of Rph3A in astrocytes but led to a decrease in the protein level of Rph3A in neurons.
The increase in Rph3A in the brain penumbra may be an endogenous protective mechanism against ischemia-reperfusion injury, which is mainly dominated by astrocytes.
研究 Rph3A 在脑缺血再灌注引起的神经元损伤中的作用。
通过 Western blot 检测半影区 Rph3A 的蛋白和 mRNA 水平。通过免疫荧光检测 Rph3A 在半影区不同细胞类型中的定位。通过 TUNEL 染色检测脑内细胞凋亡。采用转棒试验、粘取试验和 Morris 水迷宫试验进行神经行为学评价。通过 Western blot 和 ELISA 分别检测体外培养神经元和星形胶质细胞中 Rph3A 的表达和定位。
MCAO/R 模型大鼠脑半影区的 Rph3A mRNA 和蛋白水平明显升高。Rph3A 主要分布在神经元和星形胶质细胞中，MCAO/R 后明显增加。下调 Rph3A 发现，它进一步加重了大鼠 MCAO/R 后的脑梗死、神经元死亡以及行为、认知和记忆障碍。我们还发现，缺血再灌注增加了星形胶质细胞中 Rph3A 的体外蛋白水平和分泌，但导致神经元中 Rph3A 蛋白水平降低。
脑半影区 Rph3A 的增加可能是一种内源性保护机制，对抗缺血再灌注损伤，主要由星形胶质细胞主导。
