Department of Pharmacology, Chongqing Medical University, Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.
Department of Pharmacology, Chongqing Medical University, Key Laboratory of Biochemistry and Molecular Pharmacology, Chongqing 400016, China.
Biochem Pharmacol. 2024 Sep;227:116446. doi: 10.1016/j.bcp.2024.116446. Epub 2024 Jul 20.
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.
脑缺血再灌注（CIRI）是全球范围内导致死亡的主要原因之一。一种名为 ADP-核糖基化因子样蛋白 13B（ARL13B）的小分子 GTPase 在几种疾病中起着至关重要的作用。然而，ARL13B 在 CIRI 中的作用尚不清楚。我们构建了大鼠大脑中动脉闭塞/再灌注（MCAO/R）以及 PC12 细胞的氧葡萄糖剥夺/再氧合（OGD/R）模型。通过神经功能评分、TTC 染色、转棒试验、H&E 染色和尼氏染色评估 ARL13B 对 MCAO/R 的神经保护作用。使用 Western blot 和免疫荧光检测与 SHH 通路和细胞凋亡相关的蛋白表达。通过 TUNEL 检测和流式细胞术检测细胞凋亡。在脑缺血再灌注模型中，ARL13B 的表达增加。然而，ARL13B 敲低通过抑制 sonic hedgehog（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 敲低加重 CIRI 神经损伤的作用。ARL13B 减轻脑梗死和病理损伤，对 MCAO/R 起保护作用。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，同时降低促凋亡蛋白 BAX 的表达，从而减少细胞凋亡。PC12 细胞 OGD/R 模型的结果与体内结果一致。令人惊讶的是，我们证明 ARL13B 通过调节细胞周期来保护神经免受 CIRI 损伤。我们的研究结果表明，ARL13B 通过激活 SHH 依赖性通路减少细胞凋亡来保护 CIRI，表明 ARL13B 在 CIRI 发病机制中起着关键作用。
