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  

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

作　　者：孙娟 赵秀丽[1] 曾国熙[1] 张艳[2] 陈春花[2] SUN Juan;ZHAO Xiu-li;ZENG Guo-xi;ZHANG Yan;CHEN Chun-hua(Department of Neurology,Qinghai University Affiliated Hospital,Xining 810001,China;Department of Human Anatomy and Histology Embryology,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100083,China)

机构地区：[1]青海大学附属医院神经内科,西宁810001 [2]北京大学医学部基础医学院人体解剖学与组织学胚胎学系,北京100083

出　　处：《解剖学报》2021年第6期855-862,共8页Acta Anatomica Sinica

基　　金：青海省自然科学基金(2020-ZJ-948Q)。

摘　　要：目的探讨salvinorin A(SA)能否减轻大鼠蛛网膜下腔出血(SAH)后脑血管痉挛(CVS)及其机制。方法成年雄性SD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(e NOS)蛋白的表达,免疫荧光染色观察e NOS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e NOS蛋白的表达,该作用可以被PI3K抑制剂渥曼青霉素(wortmannin)和e NOS的抑制剂L-NAME所抑制;免疫荧光染色发现,e NOS表达于血管内皮细胞。结论SA能够通过PI3K/Akt/e NOS通路缓解SAH后CVS。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(e NOS)proteins were detected by Western blotting and the location of e NOS 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 e NOS proteins,which could be inhibited by PI3K inhibitor wortmannin and e NOS inhibitor L-NAME.e NOS expressed in vascular endothelial cells was detected by the immunofluorescence staining.Conclusion SA can alleviate CVS after SAH through PI3K/Akt/e NOS pathway.

关 键 词：蛛网膜下腔出血 脑血管痉挛 Salvinorin A 免疫印迹法 免疫荧光 大鼠 

分 类 号：R743.35[医药卫生—神经病学与精神病学]