Centre for Addiction and Mental Health - CAMH, Toronto, Canada; Neuroscience Program, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil.
Neuroscience Program, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brazil; Department of Pharmacology, Universidade Federal de Minas Gerais, Belo Horizonte, Brazil.
Neurosci Lett. 2021 Jun 21;756:135948. doi: 10.1016/j.neulet.2021.135948. Epub 2021 May 9.
Epilepsy is a common pathological condition that predisposes individuals to seizures, as well as cognitive and emotional dysfunctions. Different studies have demonstrated that inflammation contributes to the pathophysiology of epilepsy. Indeed, seizures change the peripheral inflammatory pattern, which, in turn, could contribute to seizures. However, the cause of the altered production of peripheral inflammatory mediators is not known. The PI3K/mTOR/GSK3β pathway is important for different physiological and pharmacological phenomena. Therefore, in the present study, we tested the hypothesis that the PI3K/mTOR/GSK3β pathway is deregulated in immune cells from patients with epilepsy and contributes to the abnormal production of inflammatory mediators.
Patients with temporal lobe epilepsy presenting hippocampal sclerosis and controls aged between 18 and 65 years-old were selected for this study. Peripheral blood was collected for the isolation of peripheral mononuclear blood cells (PBMC). Cells were pre-incubated with different PI3K, mTOR and GSK-3 inhibitors for 30 min and further stimulated with phytohaemaglutinin (PHA) or vehicle for 24 h. The supernatant was used to evaluate the production of IL-1β, IL-6, IL-10, TNF e IL-12p70.
Non-selective inhibition of PI3K, as well as inhibition of PI3Kγ and GSK-3, reduced the levels of TNF and IL-10 in PHA-stimulated cells from TLE individuals. This stimulus increased the production of IL-12p70 only in cells from TLE individuals, while the inhibition of PI3K and mTOR enhanced the production of this cytokine. On the other hand, inhibition of GSK3 reduced the PHA-induced production of IL-12p70.
Herein we demonstrated that the production of cytokines by immune cells from patients with TLE differs from non-epileptic patients. This differential regulation may be associated with the altered activity and responsiveness of intracellular molecules, such as PI3K, mTOR and GSK-3, which, in turn, might contribute to the inflammatory state that exists in epilepsy and its pathogenesis.
癫痫是一种常见的病理状态,使个体易患癫痫发作以及认知和情绪功能障碍。不同的研究表明,炎症有助于癫痫的病理生理学。事实上,癫痫改变了外周炎症模式,而反过来,这种模式可能会导致癫痫发作。然而,外周炎症介质产生改变的原因尚不清楚。PI3K/mTOR/GSK3β 途径对不同的生理和药理现象很重要。因此,在本研究中,我们测试了以下假设,即 PI3K/mTOR/GSK3β 途径在癫痫患者的免疫细胞中失调,并导致炎症介质的异常产生。
选择了患有颞叶癫痫伴海马硬化的患者和年龄在 18 至 65 岁之间的对照组参与本研究。采集外周血用于分离外周血单核细胞(PBMC)。细胞用不同的 PI3K、mTOR 和 GSK-3 抑制剂孵育 30 分钟,然后用植物血球凝集素(PHA)或载体进一步刺激 24 小时。用上清液评估 IL-1β、IL-6、IL-10、TNF 和 IL-12p70 的产生。
非选择性 PI3K 抑制以及 PI3Kγ 和 GSK-3 抑制降低了 TLE 个体 PHA 刺激细胞中 TNF 和 IL-10 的水平。这种刺激仅增加了 TLE 个体细胞中 IL-12p70 的产生,而 PI3K 和 mTOR 的抑制增强了这种细胞因子的产生。另一方面,GSK3 的抑制降低了 PHA 诱导的 IL-12p70 的产生。
在此,我们证明了 TLE 患者免疫细胞产生细胞因子的方式与非癫痫患者不同。这种差异调节可能与细胞内分子(如 PI3K、mTOR 和 GSK-3)的活性和反应性改变有关,而这些改变反过来可能有助于癫痫及其发病机制中存在的炎症状态。