Lin Juan, Lin Xinyu, Fu Yong-Juan, Li Xuran, Li Xin, Wang Yujiao, Zhao Lihong, Yu Shun, Piao Yue-Shan
Department of Pathology, Xuanwu Hospital, Capital Medical University, Beijing, 100053, People's Republic of China.
Department of Pathology, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou, 510120, People's Republic of China.
Neuropsychiatr Dis Treat. 2025 Feb 27;21:409-419. doi: 10.2147/NDT.S495339. eCollection 2025.
In this study, we evaluated the presence of tau deposition and protein expression of Reelin/GSK-3β/p-Tau signaling pathway in the hippocampus of patients with temporal lobe epilepsy (TLE).
A total of 37 cases of TLE with and without hippocampal sclerosis (HS) were selected histopathologically for our study with 5 autopsy cases as the control group. Immunohistochemistry and the histelide assay, a novel technique quantifying antigens in paraffin section, were used to confirm the distribution of protein within Reelin/GSK-3β/p-Tau signaling pathway and validate the expression of GSK-3β and AT8 (hyperphosphorylated tau) in this study.
Immunohistochemical staining for AT8 revealed punctate and filamentous positive expression in the CA1, CA2 and CA3 regions of the hippocampus with TLE under the ependyma, distributed in a band-like pattern. By contrast, the control group did not exhibit any immunopositivity. GSK-3β was strongly positive in the neuronal bodies, apical dendrites and axons in both groups of TLE, while all controls were negative. In addition, there was no significant difference in the immunohistochemical labelling of Reelin among all cases. The histelide assay indicated that the amounts of AT8 and GSK-3β were significantly increased in the two TLE groups (P < 0.05). Notably, there was a positive correlation between AT8 and GSK-3β in TLE without HS (P < 0.05).
The present data indicates that phosphorylated tau protein and GSK-3β are activated in the hippocampus of patients with TLE, and this is the first study to examine relevant proteins with the histelide assay in paraffin samples of human tissue. We consider that the regulatory network of tau protein between the two groups may be similar but not identical.
This study emphasized the Reelin/GSK-3β/p-Tau signaling pathway in TLE with a quantitative data of human tissues innovatively, revealing inspiration for mechanism exploration.
在本研究中，我们评估了颞叶癫痫（TLE）患者海马中tau蛋白沉积情况以及Reelin/GSK-3β/p-Tau信号通路的蛋白表达。
组织病理学上选取37例伴有或不伴有海马硬化（HS）的TLE患者进行研究，以5例尸检病例作为对照组。采用免疫组织化学和组织化学发光测定法（一种定量石蜡切片中抗原的新技术）来确定Reelin/GSK-3β/p-Tau信号通路中蛋白的分布，并验证本研究中GSK-3β和AT8（过度磷酸化tau蛋白）的表达。
AT8的免疫组织化学染色显示，TLE患者海马CA1、CA2和CA3区室管膜下有斑点状和丝状阳性表达，呈带状分布。相比之下，对照组未显示任何免疫阳性。两组TLE患者的神经元胞体、顶端树突和轴突中GSK-3β均呈强阳性，而所有对照组均为阴性。此外，所有病例中Reelin的免疫组织化学标记无显著差异。组织化学发光测定法表明，两个TLE组中AT8和GSK-3β的含量显著增加（P<0.05）。值得注意的是，无HS的TLE患者中AT8和GSK-3β呈正相关（P<0.05）。
目前的数据表明，TLE患者海马中磷酸化tau蛋白和GSK-3β被激活，这是首次在人体组织石蜡样本中用组织化学发光测定法检测相关蛋白的研究。我们认为两组之间tau蛋白的调控网络可能相似但不完全相同。
本研究创新性地以人体组织定量数据强调了TLE中的Reelin/GSK-3β/p-Tau信号通路，为机制探索提供了启示。
