17-烯丙基氨基-去甲氧基格尔德霉素通过HSP90/RIP3/NLRP3通路改善大鼠蛛网膜下腔出血后的微血栓形成

17-Allylamino-Demethoxygeldanamycin Ameliorate Microthrombosis Via HSP90/RIP3/NLRP3 Pathway After Subarachnoid Hemorrhage in Rats.

作者信息

Zuo Yuchun, He Tibiao, Liao Peiqiang, Zhuang Kai, Yan Xiaoxin, Liu Fei

机构信息

Department of Neurosurgery, The Third Xiangya Hospital, Central South University, Changsha, China.

Department of Anatomy, The XiangYa Medical School, Central South University, Changsha, China.

出版信息

Acta Neurochir Suppl. 2020;127:69-75. doi: 10.1007/978-3-030-04615-6_12.

DOI:10.1007/978-3-030-04615-6_12

PMID:31407066

Abstract

BACKGROUND

Subarachnoid hemorrhage (SAH) is a severe and emergent cerebrovascular disease, the prognosis of which usually very poor. Microthrombi formation highlighted with inflammation occurs early after SAH. As the main cause of DCI, microthrombosis associated with the prognosis of SAH. The aim of this study was to show HSP90 inhibitor 17-AAG effect on microthrombosis after SAH in rats.

METHODS

Ninety-five SD rats were used for the experiment. For time course study, the rats were randomly divided into five groups: sham group and SAH group with different time point (1d, 2d, 3d, 5d). Endovascular perforation method was conducted for SAH model. Neurological score, SAH grade, and mortality were measured after SAH. The samples of the left hemisphere brain were collected. The expression of HSP90 was detected by Western blot. The microthrombosis after SAH in rats' brain was detected by immunohistochemistry. For mechanism study, rats were randomly divided into three groups: sham, SAH + vehicle, and SAH +17-AAG (n = 6/group). 17-AAG was given by intraperitoneal injection (80 mg/kg) 1 h after SAH. Neurological function were measured at 24 h after SAH. The expression of RIP3, NLRP3, ASC, and IL-1β was measured by Western blot. Microthrombosis was detected by immunohistochemistry.

RESULTS

Our results showed that the HSP90 protein level increased and peaked at 2 days after SAH. Microthrombosis caused by SAH was increased in 1 day and peaked at 2 days after SAH. Administration HSP90 specific inhibitor 17-AAG reduced expression of RIP3, NLRP3, ASC, and IL-1β, reduced microthrombosis after SAH, and improved neurobehavior when compared to vehicle group.

CONCLUSIONS

17-AAG can ameliorate microthrombosis via HSP90/RIP3/NLRP3 pathway and improve neurobehavior after SAH.

摘要

背景

蛛网膜下腔出血（SAH）是一种严重的急性脑血管疾病，其预后通常很差。SAH后早期会出现伴有炎症的微血栓形成。作为迟发性脑缺血（DCI）的主要原因，微血栓形成与SAH的预后相关。本研究的目的是展示热休克蛋白90（HSP90）抑制剂17-烯丙基氨基-17-去甲氧基格尔德霉素（17-AAG）对大鼠SAH后微血栓形成的影响。

方法

95只SD大鼠用于实验。在时间进程研究中，大鼠被随机分为五组：假手术组和不同时间点（1天、2天、3天、5天）的SAH组。采用血管内穿刺法建立SAH模型。SAH后测量神经功能评分、SAH分级和死亡率。收集左半球脑样本。通过蛋白质免疫印迹法检测HSP90的表达。通过免疫组织化学法检测大鼠脑内SAH后的微血栓形成。在机制研究中，大鼠被随机分为三组：假手术组、SAH+溶媒组和SAH+17-AAG组（每组n=6）。SAH后1小时腹腔注射17-AAG（80mg/kg）。SAH后24小时测量神经功能。通过蛋白质免疫印迹法测量受体相互作用蛋白3（RIP3）、NOD样受体蛋白3（NLRP3）、凋亡相关斑点样蛋白（ASC）和白细胞介素-1β（IL-1β）的表达。通过免疫组织化学法检测微血栓形成。