科宁通过 PI3K/Akt/mTOR 通路防止自噬来保护大脑免受缺血/再灌注损伤。
Cornin protects against cerebral ischemia/reperfusion injury by preventing autophagy via the PI3K/Akt/mTOR pathway.
机构信息
Department of Pharmacology, Binzhou Medical University, Yantai, Shandong, 264003, People's Republic of China.
Department of Pharmacy, Binzhou Medical University Hospital, Binzhou, Shandong, 256603, People's Republic of China.
出版信息
BMC Pharmacol Toxicol. 2022 Oct 24;23(1):82. doi: 10.1186/s40360-022-00620-3.
BACKGROUND
Ischemia stroke is the leading cause of disability, which is a consequence of vascular occlusion. The purpose of this study is to investigate the effect of cornin which is isolated from the fruit of Verbena officinalis L, against astrocytes autophagy induced by cerebral ischemia/reperfusion (CI/R) injury in vitro and in vivo and its potential mechanism.
METHODS
Cornin at dose of 2.5, 5 and 10 mg/kg were intravenously injected to MCAO rats at 15 min after reperfusion. The infarction volume, blood-brain barrier (BBB), neurological severity score (mNSS), and autophagy related protein were used to evaluated the protective effects and potential mechanism of cornin in autophagy with or without phosphoinositide-3 kinase (PI3K)inhibitor LY294002 and mammalian target of rapamycin (mTOR) small interfering RNA (siRNA) at 24 h after CI/R injury. The potential protective effects and mechanism of cornin at concention of 10 ~ 1000 nM were also evaluated in oxygen glucose deprivation/reperfusion (OGD/R) in U87 cells.
RESULTS
The results suggest that cornin at dose of 5 or 10 mg/kg significantly reduce the cerebral infarction volume and blood-brain barrier (BBB) leakage, and improve neurological recovery in MCAO rats. Cleaved caspase-3 and Bax levels were significantly decreased, while B-cell lymphoma-2 (Bcl-2) and the apoptosis regulator ratio (Bcl-2/Bax) were markedly increased when treated with 2.5-10 mg/kg cornin. The obvious decreased expressions of glial fibrillary acidic protein (GFAP), myosin-like BCL2 interacting protein (Beclin-1) and microtubule-associated protein light chain 3 II (LC3-II) and increased of neuronal nuclei (NeuN), sequestosome-1 (p62), phosphorylated mTOR (p-mTOR), and phosphorylated Akt (p-Akt) were observed in MCAO rats treated with 10 mg/kg cornin, which was counteracted by LY294002. The expression of autophagy-related proteins with or without LY294002 and mTOR siRNA presented the similar results as in vitro in OGD/R in U87 cells.
CONCLUSIONS
These results indicate that cornin improved neurological recovery after cerebral ischemia injury by preventing astrocytes autophagy induced by CI/R via the PI3K/Akt/mTOR signaling pathway.
背景
缺血性脑卒中是导致残疾的主要原因，是血管阻塞的结果。本研究旨在探讨从马鞭草科Verbena officinalis L 的果实中分离得到的 Cornin 对体外和体内脑缺血再灌注（CI/R）损伤诱导的星形胶质细胞自噬的影响及其潜在机制。
方法
Cornin 以 2.5、5 和 10 mg/kg 的剂量于再灌注后 15 分钟通过静脉注射给予 MCAO 大鼠。采用脑梗死体积、血脑屏障（BBB）、神经严重程度评分（mNSS）和自噬相关蛋白评估 CI/R 损伤后 24 小时 Cornin 与磷酸肌醇 3 激酶（PI3K）抑制剂 LY294002 和雷帕霉素（mTOR）小干扰 RNA（siRNA）联合用药或单独用药对自噬的保护作用及其潜在机制。还评估了 Cornin 在浓度为 10~1000 nM 时对 U87 细胞氧葡萄糖剥夺/再灌注（OGD/R）的潜在保护作用和机制。
结果
结果表明，5 或 10 mg/kg 的 Cornin 可显著降低脑梗死体积和血脑屏障（BBB）渗漏，并改善 MCAO 大鼠的神经恢复。用 2.5-10 mg/kg Cornin 处理后，Cleaved caspase-3 和 Bax 水平显著降低，而 B 细胞淋巴瘤-2（Bcl-2）和凋亡调节因子比（Bcl-2/Bax）明显增加。MCAO 大鼠用 10 mg/kg Cornin 处理后，神经胶质纤维酸性蛋白（GFAP）、肌球蛋白样 BCL2 相互作用蛋白（Beclin-1）和微管相关蛋白轻链 3 II（LC3-II）的表达明显减少，神经元核（NeuN）、自噬相关蛋白 1（p62）、磷酸化 mTOR（p-mTOR）和磷酸化 Akt（p-Akt）的表达增加。LY294002 可拮抗这种作用。在 U87 细胞的 OGD/R 中，无论有无 LY294002 和 mTOR siRNA，自噬相关蛋白的表达均呈现出与体外相似的结果。
结论
这些结果表明，Cornin 通过 PI3K/Akt/mTOR 信号通路抑制 CI/R 诱导的星形胶质细胞自噬，改善了脑缺血损伤后的神经恢复。
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