Dong Jibo, Liao Yong, Wu Bihua
Department of Geriatrics, Affiliated Hospital of North Sichuan Medical College, Institute of Neurological Diseases, North Sichuan Medical College, Nanchong, China.
Ann Transl Med. 2022 Jul;10(14):795. doi: 10.21037/atm-22-2707.
There is evidence that immune inflammation plays an important role in the of epilepsy. The toll-like receptor 4 (TLR4)/nuclear factor kappa beta (NF-κB) signaling pathway is a target for the treatment of epilepsy. TAK-242 is a potent TLR4 inhibitor with neuroprotective effects. No study has examined whether TAK-242 has a protective effect on epilepsy.
Male C57BL/6 mice were randomly divided into the following 3 groups: (I) the control group; (II) the model [pentetrazol (PTZ)] group; and (III) the treatment group [PTZ + TAK-242 (3 mg/kg)], with 8 mice in each group. A mouse model of epilepsy was established via the intraperitoneal injection of PTZ (37 mg/kg). The behavioral changes of the mice were observed and scored using the Racine grading criteria. TAK-242 (3 mg/kg) was administered to establish the treatment model. The control group was intraperitoneally injected with normal saline at the same dose as the PTZ epileptogenic dose, and 3 mg/kg of normal saline was intragastrically administered. The messenger RNA (mRNA) and protein expressions of TLR4, NF-κB, and the NF-κB downstream gene tumor necrosis factor alpha (TNF-α) in the mouse brain tissues were detected by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot.
Compared to the control group, the mice in the model group showed generalized tonic convulsion and involuntary falling after PTZ modeling. The results of the hematoxylin and eosin (H&E) staining showed that the treatment group had a higher number of normal neurons than the model group, and the neuronal cell morphology in the treatment group was closer to the control group. However, the occurrence of generalized tonic convulsion and involuntary falling in the mice in the treatment group was significantly improved. Comparing the Western Blot and RT-qPCR results, we detected higher TLR4, NF-KB, TNF-α protein and mRNA expression levels in the model group than the treatment group, and there was no significant difference between the control group and the treatment group.
TAK-242 treatment ameliorates epileptic symptoms in mice, and the mechanism by which this occurs may be related to the inhibition of the TLR4/NF-κB inflammatory pathway.
有证据表明免疫炎症在癫痫发病过程中起重要作用。Toll样受体4（TLR4）/核因子κB（NF-κB）信号通路是癫痫治疗的一个靶点。TAK-242是一种具有神经保护作用的强效TLR4抑制剂。尚无研究探讨TAK-242对癫痫是否具有保护作用。
将雄性C57BL/6小鼠随机分为以下3组：（I）对照组；（II）模型[戊四氮（PTZ）]组；（III）治疗组[PTZ + TAK-242（3 mg/kg）]，每组8只小鼠。通过腹腔注射PTZ（37 mg/kg）建立癫痫小鼠模型。采用Racine分级标准观察并记录小鼠的行为变化。给予TAK-242（3 mg/kg）建立治疗模型。对照组腹腔注射与PTZ致痫剂量相同的生理盐水，并灌胃给予3 mg/kg生理盐水。采用实时定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测小鼠脑组织中TLR4、NF-κB以及NF-κB下游基因肿瘤坏死因子α（TNF-α）的信使核糖核酸（mRNA）和蛋白表达。
与对照组相比，模型组小鼠经PTZ建模后出现全身强直性惊厥和自主跌倒。苏木精-伊红（H&E）染色结果显示，治疗组正常神经元数量多于模型组，且治疗组神经元细胞形态更接近对照组。然而，治疗组小鼠全身强直性惊厥和自主跌倒的发生率显著改善。比较蛋白质免疫印迹法和RT-qPCR结果，我们检测到模型组TLR4、NF-KB、TNF-α蛋白和mRNA表达水平高于治疗组，而对照组和治疗组之间无显著差异。
TAK-242治疗可改善小鼠癫痫症状，其作用机制可能与抑制TLR4/NF-κB炎症通路有关。
