Department of Pediatrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China.
Department of Pediatrics, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, China.
Int Immunopharmacol. 2023 Aug;121:110426. doi: 10.1016/j.intimp.2023.110426. Epub 2023 Jun 8.
Neuroinflammation is a major component of epilepsy. Gut-enriched Kruppel-like factor (GKLF), a transcription factor of Kruppel-like factor family, has been reported to promote microglia activation and mediate neuroinflammation. However, the role of GKLF in epilepsy remains poorly characterized. This study focused on the function of GKLF in neuron loss and neuroinflammation in epilepsy and the molecular mechanism underlying microglia activation induced by GKLF upon lipopolysaccharides (LPS) treatment. An experimental epileptic model was induced by an intraperitoneal injection of 25 mg/kg kainic acid (KA). Lentivirus vectors (Lv) carrying Gklf CDS or short hairpin RNA targeting Gklf (shGKLF) was injected into the hippocampus, resulting in Gklf overexpression or knockdown in the hippocampus. BV-2 cells were co-infected with Lv-shGKLF or/and Lv carrying thioredoxin interacting protein (Txnip) CDS for 48 h and treated with 1 μg/mL LPS for 24 h. Results showed that GKLF enhanced KA-induced neuronal loss, pro-inflammatory cytokine secretion, activation of NOD-like receptor protein-3 (NLRP3) inflammasomes and microglia, and TXNIP expression in the hippocampus. GKLF inhibition showed negative effects on LPS-induced microglia activation, as evidenced by reduced pro-inflammatory cytokine secretion and activation of NLRP3 inflammasomes. GKLF bound to Txnip promoter and increased TXNIP expression in LPS-activated microglia. Interestingly, Txnip overexpression reversed the inhibitory effect of Gklf knockdown on microglia activation. These findings indicated that GKLF was involved in microglia activation via TXNIP. This study demonstrates the underlying mechanism of GKLF in the pathogenesis of epilepsy and uncovers that GKLF inhibition may be a therapeutic strategy for epilepsy treatment.
神经炎症是癫痫的一个主要组成部分。肠道富集的 Krüppel 样因子（GKLF），是 Krüppel 样因子家族的转录因子，据报道可促进小胶质细胞激活并介导神经炎症。然而，GKLF 在癫痫中的作用仍知之甚少。本研究主要关注 GKLF 在癫痫中神经元丢失和神经炎症中的作用，以及 GKLF 在脂多糖（LPS）处理下诱导小胶质细胞激活的分子机制。通过腹腔内注射 25mg/kg 海人酸（KA）诱导实验性癫痫模型。将携带 Gklf CDS 的慢病毒载体（Lv）或靶向 Gklf 的短发夹 RNA（shGKLF）注入海马，导致海马中 Gklf 过表达或敲低。将 BV-2 细胞与 Lv-shGKLF 或/和携带硫氧还蛋白相互作用蛋白（Txnip）CDS 的 Lv 共感染 48h，并以 1μg/mL LPS 处理 24h。结果表明，GKLF 增强了 KA 诱导的神经元丢失、促炎细胞因子分泌、NOD 样受体蛋白-3（NLRP3）炎症小体和小胶质细胞的激活以及 TXNIP 在海马中的表达。GKLF 抑制对 LPS 诱导的小胶质细胞激活具有负向作用，表现为促炎细胞因子分泌减少和 NLRP3 炎症小体激活减少。GKLF 结合到 Txnip 启动子并增加 LPS 激活的小胶质细胞中的 TXNIP 表达。有趣的是，Txnip 过表达逆转了 Gklf 敲低对小胶质细胞激活的抑制作用。这些发现表明 GKLF 通过 TXNIP 参与小胶质细胞激活。本研究阐明了 GKLF 在癫痫发病机制中的潜在机制，并揭示了 GKLF 抑制可能是治疗癫痫的一种策略。
