Wu Zhenggang, Liu Jing, Yin Deju, Huang Jing, Huang Yujing, Wang Pengfei
Department of Neurology, The Affiliated Taizhou People's Hospital of Nanjing Medical University, Taizhou, Jiangsu, China.
Research and Development Department, Jiangsu Zhiyu Medical Technology Co., LTD, Taizhou, Jiangsu, China.
Organogenesis. 2025 Dec;21(1):2519607. doi: 10.1080/15476278.2025.2519607. Epub 2025 Jun 26.
Epilepsy is a common disease of the nervous system. Recent advances in epigenetics have revealed DNA methylation as a key mechanism in epilepsy pathogenesis, particularly through dysregulation of GABAergic signaling. Baicalein has been shown to have anticonvulsant and neuroprotective effects. However, its epigenetic regulatory effects on GABA receptor function remain unexplored.
The status epilepticus (SE) model was induced by lithium chloride-pilocarpine (LiCl-PILO) in Sprague-Dawley (SD) rats. The rats were divided into control group, epileptic SE group and baicalein intervention group. Morris water maze (MWM) test, Nissl staining, immunofluorescence and enzyme-linked immunosorbent assay (ELISA) were used to detect cognitive functions and neuronal damage. Online sites, chromatin immunoprecipitation (ChIP) and western blotting were used to identify DNA methyltransferase 1 (DNMT1)-mediated methylation of gamma-aminobutyric acid type A receptor subunit delta (GABRD) promoter region.
Baicalein treatment significantly prolonged the latency of SE onset and seizure onset, and improved the development of epilepsy. Meanwhile, baicalein improved the cognitive impairment in rats induced by LiCl-PILO. After treatment with baicalein, a sustained elevation in the number of neurons and NeuN levels was observed, along with a decrease in the contents of tumor necrosis factor -alpha (TNF-α), interleukin-1β (IL-1β), and ionized calcium-binding adapter molecule 1 (Iba-1) in the hippocampus. Mechanistically, baicalein interacted with DNMT1 to suppress GABRD promoter region methylation, thus increasing GABRD protein level in the hippocampus of rats induced by LiCl-PILO.
This study identifies DNMT1/GABRD axis as a novel epigenetic target for epilepsy intervention. Baicalein's ability to enhance tonic inhibition through demethylation of GABRD provides a groundbreaking strategy for drug-resistant epilepsy.
癫痫是一种常见的神经系统疾病。表观遗传学的最新进展揭示了DNA甲基化是癫痫发病机制中的关键机制，特别是通过γ-氨基丁酸能信号传导的失调。黄芩素已被证明具有抗惊厥和神经保护作用。然而，其对GABA受体功能的表观遗传调控作用仍未被探索。
采用氯化锂-匹罗卡品（LiCl-PILO）诱导Sprague-Dawley（SD）大鼠癫痫持续状态（SE）模型。将大鼠分为对照组、癫痫SE组和黄芩素干预组。采用Morris水迷宫（MWM）试验、尼氏染色、免疫荧光和酶联免疫吸附测定（ELISA）检测认知功能和神经元损伤。利用在线网站、染色质免疫沉淀（ChIP）和蛋白质印迹法鉴定DNA甲基转移酶1（DNMT1）介导的γ-氨基丁酸A型受体δ亚基（GABRD）启动子区域的甲基化。
黄芩素治疗显著延长了SE发作和癫痫发作的潜伏期，并改善了癫痫的发展。同时，黄芩素改善了LiCl-PILO诱导的大鼠认知障碍。黄芩素治疗后，观察到神经元数量和NeuN水平持续升高，同时海马中肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）和离子钙结合衔接分子1（Iba-1）的含量降低。机制上，黄芩素与DNMT1相互作用以抑制GABRD启动子区域甲基化，从而增加LiCl-PILO诱导的大鼠海马中GABRD蛋白水平。
本研究确定DNMT1/GABRD轴是癫痫干预的一个新的表观遗传靶点。黄芩素通过去甲基化GABRD增强强直抑制的能力为耐药性癫痫提供了一种开创性的策略。
