神经炎症是出血性中风后的关键致病事件。内质网(ER)应激诱导的细胞凋亡和核苷酸结合域,富含亮氨酸的重复,和含pyrin结构域的蛋白3(NLRP3)相关的焦亡可以促进神经炎症反应的升级,导致脑损伤增加。G蛋白偶联雌激素受体1(GPER1),作为最广泛特征的脑源性雌激素,据报道会触发神经保护作用。然而,GPER1激活的抗凋亡和抗发热作用及其潜在机制尚未完全阐明.我们通过血管内穿孔建立了实验性SAH模型。SAH后1小时静脉内施用GPER1选择性激动剂G1。对于机械勘探,单磷酸腺苷活化蛋白激酶(AMPK)的选择性抑制剂,dorsomorphin,在SAH诱导前30分钟通过侧脑室注射给药。SAH后评估包括SAH等级,短期和长期的神经系统结果,脑水肿,脑血流量,透射电子显微镜(TEM),蛋白质印迹(WB),ELISA,TUNEL染色,Fluoro-JadeC染色(FJC),和免疫荧光染色。观察到GPER1的表达在SAH后6小时升高,并在24小时达到峰值,主要与神经元共定位。G1治疗后显着改善SAH小鼠的短期和长期神经功能缺损,以及抑制神经元内质网应激相关凋亡蛋白的表达(即,CHOP,GRP78,caspase-12,cleavedcaspase-3,Bax,Bcl2)和焦亡相关蛋白(即,NLRP3,ASC,裂解的Caspase-1)。此外,我们的研究表明,用dorsomorphin抑制AMPK减弱了G1的神经保护作用。这伴随着与ER应激诱导的细胞凋亡和焦亡相关的分子途径的改变。本文的这些数据阐明了GPER1通过以AMPK依赖性方式减轻神经炎症而发挥神经保护作用。调节神经元内质网应激相关的凋亡和焦亡。通过激活GPER1增强抗凋亡和抗发热作用可能是SAH患者的有效治疗策略。
Neuroinflammation is a critical pathogenic event following hemorrhagic stroke. Endoplasmic reticulum (ER) stress-induced apoptosis and nucleotide-binding domain, leucine-rich repeat, and pyrin domain-containing protein 3(NLRP3)-associated pyroptosis can contribute to the escalation of neuroinflammatory responses, leading to increased brain damage. G protein-coupled estrogen receptor 1(
GPER1), as the most extensively characterized brain-derived estrogen, was reported to trigger neuroprotective effects. However, the anti-apoptotic and anti-pyroptotic effect of
GPER1 activation and the underlying mechanism has not been fully elucidated. We established the experimental SAH model by intravascular perforation. The
GPER1 selective agonist G1 was intravenously administered 1 h following SAH. For mechanistic exploration, the selective inhibitor of adenosine monophosphate-activated protein kinase (AMPK), dorsomorphin, was administered via intracerebroventricular injection 30 min prior to SAH induction. Post-SAH assessments included SAH grade, the short-term and long-term neurological outcomes, brain edema, cerebral blood flow, transmission electron microscopy (TEM), western blot (WB), ELISA, TUNEL staining, Fluoro-Jade C staining (FJC), and immunofluorescence staining. The expression of GPER1 was observed to elevate at 6 h and peaked at 24 h subsequent to SAH, predominantly co-localized with neurons. Post-treatment with G1 markedly ameliorated both the short-term and long-term neurological deficits of SAH mouse, as well as inhibiting the expression of neuronal ER stress-associated apoptotic proteins (i.e., CHOP, GRP78, Caspase-12, Cleaved Caspase-3, Bax, Bcl2) and pyroptosis-associated proteins (i.e., NLRP3, ASC, Cleaved Caspase-1). Additionally, our research revealed that inhibition of AMPK with dorsomorphin attenuated the neuroprotective effects of G1. This was accompanied by modifications in the molecular pathways associated with ER stress-induced apoptosis and pyroptosis. These data herein elucidated that
GPER1 exerted neuroprotective effects by mitigating neuroinflammation in an AMPK-dependent manner, which modulates neuronal ER stress-associated apoptosis and pyroptosis. Boosting the anti-apoptotic and anti-pyroptotic effect by activating
GPER1 may be an efficient treatment strategy for SAH patients.