Abstract：
Cerebral ischemia/reperfusion (CIRI) is a leading cause of death worldwide. A small GTPase known as ADP-ribosylation factor-like protein 13B (ARL13B) is essential in several illnesses. The role of ARL13B in CIRI remains unknown, though. A middle cerebral artery occlusion/reperfusion (MCAO/R) in rats as well as an oxygen-glucose deprivation/reoxygenation (OGD/R) models in PC12 cells were constructed. The neuroprotective effects of ARL13B against MCAO/R were evaluated using neurological scores, TTC staining, rotarod testing, H&E staining, and Nissl staining. To detect the expression of proteins associated with the SHH pathway and apoptosis, western blotting and immunofluorescence were employed. Apoptosis was detected using TUNEL assays and flow cytometry. There was increased expression of ARL13B in cerebral ischemia/reperfusion models. However, ARL13B knockdown aggravated CIRI nerve injury by inhibiting the sonic hedgehog (SHH) pathway. In addition, the use of SHH pathway agonist (SAG) can increased ARL13B expression, reverse the effects of ARL13B knockdown exacerbating CIRI nerve injury. ARL13B alleviated cerebral infarction and pathological injury and played a protective role against MCAO/R. Furthermore, ARL13B significantly increased the expression of SHH pathway-related proteins and the anti-apoptotic protein BCL-2, while decreased the expression of pro-apoptotic protein BAX, thus reducing apoptosis. The results from the OGD/R model in PC12 cells were consistent with those obtained in vivo. Surprisingly, we demonstrated that ARL13B regulates the cell cycle to protect against CIRI nerve injury. Our findings indicate that ARL13B protects against CIRI by reducing apoptosis through SHH-dependent pathway activation, and suggest that ARL13B plays a crucial role in CIRI pathogenesis.
摘要：
脑缺血/再灌注(CIRI)是世界范围内的主要死亡原因。一种称为ADP-核糖基化因子样蛋白13B（ARL13B）的小GTP酶在几种疾病中必不可少。ARL13B在CIRI中的作用仍然未知，不过。建立大鼠大脑中动脉阻塞/再灌注（MCAO/R）和PC12细胞氧糖剥夺/复氧（OGD/R）模型。使用神经评分评估ARL13B对MCAO/R的神经保护作用，TTC染色,旋转杆测试,H&E染色,和Nissl染色。检测SHH通路与细胞凋亡相关蛋白的表达,采用蛋白质印迹和免疫荧光。使用TUNEL测定和流式细胞术检测细胞凋亡。脑缺血/再灌注模型中ARL13B的表达增加。然而,ARL13B敲低可通过抑制声波刺猬（SHH）通路加重CIRI神经损伤。此外,使用SHH通路激动剂（SAG）可以增加ARL13B的表达，逆转ARL13B敲除exerbatingCIRI神经损伤的作用。ARL13B减轻脑梗死和病理损伤，对MCAO/R具有保护作用。此外，ARL13B显著增加SHH通路相关蛋白和抗凋亡蛋白BCL-2的表达,而降低促凋亡蛋白BAX的表达,从而减少细胞凋亡。PC12细胞中OGD/R模型的结果与体内获得的结果一致。令人惊讶的是,我们证明ARL13B调节细胞周期以保护抗CIRI神经损伤。我们的发现表明，ARL13B通过SHH依赖性途径激活减少细胞凋亡来保护抗CIRI，提示ARL13B在CIRI发病机制中起着至关重要的作用。
