Department of Neurosurgery, Henan Provincial People's Hospital, (People's Hospital of Zhengzhou University), Zhengzhou, 450003, China.
Department of Physiology and Pharmacology, Loma Linda University, Loma Linda, CA, 92350, USA.
Neurotherapeutics. 2023 Apr;20(3):822-837. doi: 10.1007/s13311-022-01330-8. Epub 2022 Dec 8.
Estrogen-related receptors (ERRs) were shown to play an important role in the regulation of free radical-mediated pathology. This study aimed to investigate the neuroprotective effect of ERRγ activation against early brain injury (EBI) after subarachnoid hemorrhage (SAH) and the potential underlying mechanisms. In a rat model of SAH, the time course of ERRs and SIRT3 and the effects of ERRγ activation were investigated. ERRγ agonist DY131, selective inhibitor GSK5182, or SIRT3 selective inhibitor 3-TYP were administered intracerebroventricularly (icv) in the rat model of SAH. The use of 3-TYP was for validating SIRT3 as the downstream signaling of ERRγ activation. Post-SAH assessments included SAH grade, neurological score, Western blot, Nissl staining, and immunofluorescence staining in rats. In an vitro study, the ERRγ agonist DY131 and ERRγ siRNA were administered to primary cortical neurons stimulated by Hb, after which cell viability and neuronal deaths were accessed. Lastly, the brain ERRγ levels and neuronal death were accessed in SAH patients. We found that brain ERRγ expressions were significantly increased, but the expression of SIRT3 dramatically decreased after SAH in rats. In the brains of SAH rats, ERRγ was expressed primarily in neurons, astrocytes, and microglia. The activation of ERRγ with DY131 significantly improved the short-term and long-term neurological deficits, accompanied by reductions in oxidative stress and neuronal apoptosis at 24 h after SAH in rats. DY131 treatment significantly increased the expressions of PGC-1α, SIRT3, and Bcl-2 while downregulating the expressions of 4-HNE and Bax. ERRγ antagonist GSK5182 and SIRT3 inhibitor 3-TYP abolished the neuroprotective effects of ERRγ activation in the SAH rats. An in vitro study showed that Hb stimulation significantly increased intracellular oxidative stress in primary cortical neurons, and DY131 reduced such elevations. Primary cortical neurons transfected with the ERRγ siRNA exhibited notable apoptosis and abolished the protective effect of DY131. The examination of SAH patients' brain samples revealed increases in ERRγ expressions and neuronal apoptosis marker CC3. We concluded that ERRγ activation with DY131 ameliorated oxidative stress and neuronal apoptosis after the experimental SAH. The effects were, at least in part, through the ERRγ/PGC-1α/SIRT3 signaling pathway. ERRγ may serve as a novel therapeutic target to ameliorate EBI after SAH.
雌激素相关受体(ERRs)在调节自由基介导的病理学中发挥着重要作用。本研究旨在探讨 ERRγ 激活对蛛网膜下腔出血(SAH)后早期脑损伤(EBI)的神经保护作用及其潜在机制。在 SAH 大鼠模型中,研究了 ERRs 和 SIRT3 的时程变化以及 ERRγ 激活的作用。在 SAH 大鼠模型中,通过脑室内(icv)给予 ERRγ 激动剂 DY131、选择性抑制剂 GSK5182 或 SIRT3 选择性抑制剂 3-TYP。使用 3-TYP 是为了验证 SIRT3 是 ERRγ 激活的下游信号。SAH 后评估包括 SAH 分级、神经评分、Western blot、尼氏染色和免疫荧光染色。在体外研究中,用 Hb 刺激原代皮质神经元后,给予 ERRγ 激动剂 DY131 和 ERRγ siRNA,然后评估细胞活力和神经元死亡。最后,在 SAH 患者中评估大脑 ERRγ 水平和神经元死亡。我们发现,在 SAH 后,大鼠大脑中 ERRγ 的表达明显增加,但 SIRT3 的表达显著下降。在 SAH 大鼠的大脑中,ERRγ 主要表达在神经元、星形胶质细胞和小胶质细胞中。用 DY131 激活 ERRγ 可显著改善大鼠 SAH 后 24 小时的短期和长期神经功能缺损,同时降低氧化应激和神经元凋亡。DY131 治疗可显著增加 PGC-1α、SIRT3 和 Bcl-2 的表达,同时下调 4-HNE 和 Bax 的表达。ERRγ 拮抗剂 GSK5182 和 SIRT3 抑制剂 3-TYP 消除了 ERRγ 激活在 SAH 大鼠中的神经保护作用。体外研究表明,Hb 刺激显著增加了原代皮质神经元的细胞内氧化应激,而 DY131 降低了这种升高。转染 ERRγ siRNA 的原代皮质神经元表现出明显的凋亡,并消除了 DY131 的保护作用。对 SAH 患者脑样本的检查显示,ERRγ 的表达和神经元凋亡标志物 CC3 增加。我们得出结论,用 DY131 激活 ERRγ 可改善实验性 SAH 后的氧化应激和神经元凋亡。这些作用至少部分是通过 ERRγ/PGC-1α/SIRT3 信号通路实现的。ERRγ 可能成为改善 SAH 后 EBI 的新治疗靶点。