司库奇尤单抗通过 IL-17RA/AKT/ERK1/2 通路减轻脓毒症大鼠模型的氧化应激和神经元凋亡，从而缓解认知障碍。

Secukinumab alleviates cognitive impairment by attenuating oxidative stress and neuronal apoptosis via the IL-17RA/AKT/ERK1/2 pathway in a rat model of sepsis.

机构信息

Department of Emergency, Huashan Hospital, Fudan University, 12 Middle Wulumuqi, Shanghai, China.

出版信息

Exp Neurol. 2023 Jan;359:114263. doi: 10.1016/j.expneurol.2022.114263. Epub 2022 Nov 4.

DOI:10.1016/j.expneurol.2022.114263

PMID:36336029

Abstract

BACKGROUND

Septic-associated encephalopathy (SAE) is a critical manifestation of sepsis that leads to long-term cognitive impairment. Interleukin (IL)-17A has been shown to mediate neuronal apoptosis in central nervous system diseases, while oxidative stress has been found to have a detrimental effect in SAE. However, the relationship between IL-17A and oxidative stress in SAE remains unclear. This study aimed to investigate the effects of secukinumab on alleviating cognitive impairment in a rat model of sepsis, as well as examine its underlying molecular mechanism of action.

METHODS

A total of 282 male 8-week-old Sprague-Dawley rats were randomly subjected to cecal ligation and puncture (CLP) or sham treatment followed by volume resuscitation immediately after surgery. Secukinumab was administered intranasally 1 h post-CLP. Rats were given the p-ERK activator ceramide C6 intracerebroventricularly (i.c.v) 24 h before CLP surgery. Recombinant rIL-17A was administered i.c.v. at 0 h in naive rats, followed by intraperitoneal injection of the AKT inhibitor GDC0068 1 h post-rIL-17A injection. Clinical scores, body weight, and survival rate were assessed. In addition, immunofluorescence staining, neurobehavioral tests, Nissl staining, and western blotting were performed. Cognitive function was assessed 15-20 days post-CLP using the Morris water maze test.

RESULTS

IL-17A and IL-17RA protein expression levels in the rat hippocampus increased and peaked 24 h post-CLP. Furthermore, IL-17RA was found to be expressed in neurons. The survival rate after CLP was 50%. Following CLP, an increased clinical score and significant decrease in body weight were observed. However, treatment with secukinumab led to a decrease in the clinical score of rats 24 h post-CLP. CLP resulted in spatial and memory impairment and anxiety-like behaviors in rats, while secukinumab treatment significantly alleviated cognitive impairment compared to the CLP group (p < 0.05). In addition, oxidative stress and neuronal apoptosis were found to be increased in the CLP group, while secukinumab significantly reduced oxidative stress and neuronal apoptosis in the hippocampus following CLP. Furthermore, secukinumab treatment led to a significant decrease in the protein expression levels of p-AKT, p-ERK1/2, Romo1, and Bax, together with increased Bcl-2 protein expression. Finally, treatment with ceramide C6 and GDC0068 abolished the neuroprotective effects of secukinumab post-CLP.

CONCLUSION

Our results demonstrated that secukinumab attenuated oxidative stress and neuronal apoptosis and partially ameliorated cognitive impairment via the IL-17RA/AKT/ERK1/2 pathway in a rat model of sepsis. Thus, secukinumab may be a potential therapeutic strategy for septic patients.

摘要

背景

脓毒症相关性脑病（SAE）是脓毒症的一种严重表现，可导致长期认知障碍。白细胞介素（IL）-17A 已被证明可介导中枢神经系统疾病中的神经元凋亡，而氧化应激已被发现对 SAE 具有有害影响。然而，SAE 中 IL-17A 和氧化应激之间的关系仍不清楚。本研究旨在探讨 secukinumab 对减轻脓毒症大鼠模型认知障碍的影响，并研究其潜在的作用机制。

方法

将 282 只雄性 8 周龄 Sprague-Dawley 大鼠随机分为盲肠结扎穿孔（CLP）组或假手术组，术后立即进行容量复苏。CLP 后 1h 经鼻给予 secukinumab。在 CLP 术前 24h，将 p-ERK 激活剂神经酰胺 C6 鞘内给药。在幼稚大鼠中，于 0h 经脑室给予重组 rIL-17A，1h 后给予 AKT 抑制剂 GDC0068 腹腔注射。评估临床评分、体重和存活率。此外，还进行了免疫荧光染色、神经行为测试、尼氏染色和 Western blot 分析。CLP 后 15-20 天，采用 Morris 水迷宫测试评估认知功能。

结果

CLP 后大鼠海马 IL-17A 和 IL-17RA 蛋白表达水平升高，并在 24h 达到峰值。此外，IL-17RA 被发现存在于神经元中。CLP 后大鼠的存活率为 50%。CLP 后，临床评分升高，体重明显下降。然而，secukinumab 治疗可降低 CLP 后大鼠的临床评分。CLP 导致大鼠空间和记忆障碍以及焦虑样行为，而 secukinumab 治疗与 CLP 组相比显著减轻了认知障碍（p<0.05）。此外，CLP 组氧化应激和神经元凋亡增加，而 secukinumab 显著降低了 CLP 后海马的氧化应激和神经元凋亡。此外，secukinumab 治疗可显著降低 p-AKT、p-ERK1/2、Romo1 和 Bax 的蛋白表达水平，同时增加 Bcl-2 蛋白的表达。最后，鞘内给予神经酰胺 C6 和 GDC0068 可消除 secukinumab 在 CLP 后的神经保护作用。