Salvinorin A通过磷脂酰肌醇-3激酶/蛋白激酶B/ 内皮型一氧化氮合酶通路减轻蛛网膜下腔出血后脑血管痉挛

孙娟; 张艳; 陈春花

doi:10.16098/j.issn.0529-1356.2021.06.003

PDF(6828 KB)

解剖学报 ›› 2021, Vol. 52 ›› Issue (6) : 855-862. DOI: 10.16098/j.issn.0529-1356.2021.06.003

神经生物学

Salvinorin A通过磷脂酰肌醇-3激酶/蛋白激酶B/ 内皮型一氧化氮合酶通路减轻蛛网膜下腔出血后脑血管痉挛

孙娟¹ 赵秀丽 ¹曾国熙 ¹ 张艳^2* 陈春花^2*

作者信息 +

Salvinorin A alleviating cerebral vasospasm after subarachnoid hemorrhage through phosphatidylinositol 3-kinase/protein kinase B/endothelial nitric oxide synthase pathway

SUN Juan¹ ZHAO Xiu-li¹ZENG Guo-xi¹ ZHANG Yan^2* CHEN Chun-hua^2*

Author information +

文章历史 +

摘要

目的探讨salvinD大鼠（n=97），采用颈内动脉刺破法建立大鼠SAH模型，随机分为假手术组（sham）、SAH模型组（SAH）、溶剂对照组（SAH+DMSO）和给药组（SAH+SA），SA及溶剂DMSO于SAH模型后24 h、48 h及72 h用生理盐水稀释后腹腔注射；SAH后72 h检测大鼠神经功能学评分，HE染色观察颈内动脉的血管内径和血管壁厚度，内皮素-1（ET-1）ELISA试剂盒和一氧化氮（NO）试剂盒检测Willis环血管上ET-1浓度和NO含量，Western blotting检测磷酸化PI3K(p-PI3K）、PI3K、磷酸化Akt（p-Akt）、Akt及内皮型一氧化氮合酶（eNOS）蛋白的表达，免疫荧光染色观察eNOS蛋白的表达位置。结果 SAH后72 h，SA能够升高SAH后的神经功能水平，增加SAH后血管内径，降低血管壁厚度，SA降低SAH后Willis环血管上ET-1浓度并升高NO含量；SA能够升高p-PI3K/PI3K、p-Akt/Akt及eNOS蛋白的表达，该作用可以被PI3K抑制剂渥曼青霉素（wortmannin）和eNOS的抑制剂L-NAME所抑制；免疫荧光染色发现，eNOS表达于血管内皮细胞。结论 SA能够通过PI3K/Akt/eNOS通路缓解SAH后CVS。

Abstract

Objective To investigate the effect of salvinorin A (SA) on alleviating cerebral vasospasm (CVS) after subarachnoid hemorrhage (SAH). Methods The SAH models were established by endovascular perforation method . Adult male SD rats (n=97) were randomly divided into the sham group (sham), SAH model group (SAH), control group (SAH+DMSO) and drug administration group (SAH+SA). SA and DMSO were diluted with saline, and injected intraperitoneally at hour 24, hour 48 and hour 72 after SAH. At hour 72 after SAH, the neurological score was evaluated. The diameter and wall thickness of the internal carotid artery were observed through HE staining. Endothelin 1 (ET-1) ELISA kit and nitric oxide (NO) kit were used to observe the ET-1 concentration and NO content on the blood vessels of Willis circle. The expression of phosphorglated PI3K(p-PI3K), PI3K, phosphorylated Akt(p-Akt), Akt and endothelial nitric oxide synthase (eNOS) proteins were detected by Western blotting and the location of eNOS protein was observed by immunofluorescent staining. Results At hour 72 after SAH, SA could increase the neurological score, increase the vessel diameter and reduce the wall thickness of internal carotid artery. SA could reduce the ET-1 concentration and increase NO content in the blood vessels of Willis circle at hour 72 after SAH. SA could increase the ratio of p-PI3K/PI3K, p-Akt/Akt and the expression of eNOS proteins, which could be inhibited by PI3K inhibitor wortmannin and eNOS inhibitor L-NAME. eNOS expressed in vascular endothelial cells was detected by the immunofluorescence staining. Conclusion SA can alleviate CVS after SAH through PI3K/Akt/eNOS pathway.

导出引用

孙娟赵秀丽曾国熙张艳陈春花. Salvinorin A通过磷脂酰肌醇-3激酶/蛋白激酶B/ 内皮型一氧化氮合酶通路减轻蛛网膜下腔出血后脑血管痉挛[J]. 解剖学报. 2021, 52(6): 855-862 https://doi.org/10.16098/j.issn.0529-1356.2021.06.003

SUN Juan ZHAO Xiu-li ZENG Guo-xi ZHANG Yan CHEN Chun-hua. Salvinorin A alleviating cerebral vasospasm after subarachnoid hemorrhage through phosphatidylinositol 3-kinase/protein kinase B/endothelial nitric oxide synthase pathway[J]. Acta Anatomica Sinica. 2021, 52(6): 855-862 https://doi.org/10.16098/j.issn.0529-1356.2021.06.003

中图分类号： R743.35

参考文献

［1］ Liu SQ, Zhang Y, Sun J, et al. Progress of mechanism and therepy on cerebral vasospasm after subarachnoid hemorrhage［J］. Acta Anatomica Sinca, 2019, 50（4）: 537-542. (in Chinese)

刘思齐，张艳，孙娟，等. 蛛网膜下腔出血后继发性脑血管痉挛的研究进展［J］.解剖学报, 2019, 50（4）: 537-542.

［2］ Li G, Wang QS, Lin TT. Alterations in the expression of protease-activated receptor 1 and tumor necrosis factorα in the basilar artery of rats following a subarachnoid hemorrhage［J］. Exp Ther Med, 2016, 11(3): 717-722.

［3］ Xu T, Zhang WG, Sun J, et al. Protective effects of thrombomodulin on microvascular permeability after subarachnoid hemorrhage in mouse model［J］. Neuroscience, 2015; 299:18-27.

［4］ He J, Liu M, Liu Z, et al. Recombinant osteopontin attenuates experimental cerebral vasospasm following subarachnoid hemorrhage in rats through an anti-apoptotic mechanism［J］. Brain Res, 2015, 1611: 74-83.

［5］ Chang CZ, Wu SC, Chang CM, et al. Arctigenin, a potent ingredient of arctium lappa l, induces endothelial nitric oxide synthase and attenuates subarachnoid hemorrhage-induced vasospasm through PI3K/Akt pathway in a rat model［J］. Biomed Res Int, 2015, 2015: 490209.

［6］ Peng XQ, Damarla M, Skirball J, et al. Protective role of PI3-kinase/Akt/eNOS signaling in mechanical stress through inhibition of p38 mitogen-activated protein kinase in mouse lung［J］. Acta Pharmacol Sin, 2010, 31(2): 175-183.

［7］ Kivell B, Uzelac Z, Sundaramurthy S, et al. Salvinorin A regulates dopamine transporter function via a kappa opioid receptor and ERK1/2-dependent mechanism［J］. Neuropharmacology, 2014, 86: 228-240.

［8］ Addy PH. Acute and post-acute behavioral and psychological effects of salvinorin A in humans ［J］. Psychopharmacology (Berl), 2012, 220(1): 195-204.

［9］ Sun J, Yang XM, Zhang Y, et al. Salvinorin A attenuates early brain injury through PI3K/Akt pathway after subarachnoid hemorrhage in rat［J］. Brain Res, 2019,1719:64-70.

［10］ Su D, Riley J, Kiessling WJ, et al. Salvinorin A produces cerebrovasodilation through activation of nitric oxide synthase, kappa receptor, and adenosine triphosphate-sensitive potassium channel［J］. Anesthesiology, 2011, 114(2): 374-379.

［11］ Sun J, Zhang Y, Lu JF, et al. Salvinorin A ameliorates cerebral vasospasm through activation of endothelial nitric oxide synthase in a rat model of subarachnoid hemorrhage［J］. Microcirculation, 2018, 25(3):e12442.

［12］ Hosseini M, Sadeghnia HR, Salehabadi S, et al. The effect of L-arginine and L-NAME on pentylenetetrazole induced seizures in ovariectomized rats, an in vivo study［J］. Seizure, 2009, 18(10): 695-698.

［13］ Chang B, Sang L, Wang Y, et al. The role of FoxO4 in the relationship between alcohol-induced intestinal barrier dysfunction and liver injury［J］. Int J Mol Med, 2013, 31(3): 569-576.

［14］ Bederson JB, Germano IM, Guarino L. Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat［J］. Stroke, 1995, 26(6): 1086-1092.

［15］ Garcia JH, Wagner S, Liu KF, et al. Neurological deficit and extent of neuronal necrosis attributable to middle cerebral artery occlusion in rats. Statistical validation［J］. Stroke, 1995, 26(4): 627-635.

［16］ Hasegawa S, Hasegawa Y, Miura M. Current therapeutic durgs against cerebral vasospsm after subarachnoid hemorrhage: a comprehensive review of basic and clinial studies［J］. Curr Drug Deliv, 2017, 14(6): 843-852.

［17］ Nishizawa S, Laher Ⅰ. Signaling mechanisms in cerebral vasospasm［J］. Trends Cardiovasc Med, 2005, 15(1): 24-34.

［18］ Cho HG, Shin HK, Shin YW, et al. Role of nitric oxide in the CBF autoregulation during acute stage after subarachnoid haemorrhage in rat pial artery［J］. Fundam Clin Pharmacol,2003, 17(5): 563-573

［19］ Pluta RM. Delayed cerebral vasospasm and nitric oxide: review, new hypothesis, and proposed treatment［J］. Pharmacol Ther, 2005, 105(1): 23-56.

［20］ Dietrich HH, Dacey Jr RG. Molecular keys to the problems of cerebral vasospasm［J］. Neurosurgery, 2000, 46(3): 517-530.

［21］ Zhu J, Song W, Li L, et al. Endothelial nitric oxide synthase:a potential therapeutic target for cerebrovascular diseases［J］. Mol Brain,2016,9(1):30-37.

［22］ Blanes MG, Oubaha M, Rautureau Y, et al. Phosphorylation of tyrosine 801 of vascular endothelial growth factor receptor-2 is necessary for Akt-dependent endothelial nitric-oxide synthase activation and nitric oxide release from endothelial cells［J］. J Biol Chem, 2007, 282(14): 10660-0669.