Table_1_Novel insights into STIM1's role in store-operated calcium entry and its implications for T-cell mediated inflammation in trigeminal neuralgia.DOCX

This investigation aims to elucidate the novel role of Stromal Interaction Molecule 1 (STIM1) in modulating store-operated calcium entry (SOCE) and its subsequent impact on inflammatory cytokine release in T lymphocytes, thereby advancing our understanding of trigeminal neuralgia (TN) pathogenesis. Employing the Gene Expression Omnibus (GEO) database, we extracted microarray data pertinent to TN to identify differentially expressed genes (DEGs). A subsequent comparison with SOCE-related genes from the Genecards database helped pinpoint potential target genes. The STRING database facilitated protein-protein interaction (PPI) analysis to spotlight STIM1 as a gene of interest in TN. Through histological staining, transmission electron microscopy (TEM), and behavioral assessments, we probed STIM1's pathological effects on TN in rat models. Additionally, we examined STIM1's influence on the SOCE pathway in trigeminal ganglion cells using techniques like calcium content measurement, patch clamp electrophysiology, and STIM1- ORAI1 co-localization studies. Changes in the expression of inflammatory markers (TNF-α, IL-1β, IL-6) in T cells were quantified using Western blot (WB) and enzyme-linked immunosorbent assay (ELISA) in vitro, while immunohistochemistry and flow cytometry were applied in vivo to assess these cytokines and T cell count alterations. Our bioinformatic approach highlighted STIM1's significant overexpression in TN patients, underscoring its pivotal role in TN's etiology and progression. Experimental findings from both in vitro and in vivo studies corroborated STIM1's regulatory influence on the SOCE pathway. Furthermore, STIM1 was shown to mediate SOCE-induced inflammatory cytokine release in T lymphocytes, a critical factor in TN development. Supportive evidence from histological, ultrastructural, and behavioral analyses reinforced the link between STIM1-mediated SOCE and T lymphocyte-driven inflammation in TN pathogenesis. This study presents novel evidence that STIM1 is a key regulator of SOCE and inflammatory cytokine release in T lymphocytes, contributing significantly to the pathogenesis of trigeminal neuralgia. Our findings not only deepen the understanding of TN's molecular underpinnings but also potentially open new avenues for targeted therapeutic strategies.

本研究旨在阐明基质相互作用分子1（Stromal Interaction Molecule 1，STIM1）在调控钙池操纵性钙内流（store-operated calcium entry，SOCE）中的新功能，及其对T淋巴细胞（T lymphocytes）中炎症细胞因子释放的后续影响，进而深化对三叉神经痛（trigeminal neuralgia，TN）发病机制的认知。本研究依托基因表达综合数据库（Gene Expression Omnibus，GEO），提取与TN相关的微阵列数据以筛选差异表达基因（differentially expressed genes，DEGs）；通过与基因卡数据库（Genecards）中的SOCE相关基因进行比对，进一步锁定潜在靶基因。借助STRING数据库开展蛋白质相互作用（protein-protein interaction，PPI）分析，最终将STIM1确定为TN研究的目标基因。通过组织化学染色、透射电子显微镜（transmission electron microscopy，TEM）及行为学评估，我们在大鼠模型中探究了STIM1对TN的病理效应。此外，我们采用钙含量检测、膜片钳电生理技术及STIM1-ORAI1共定位分析等方法，检测了STIM1对三叉神经节细胞中SOCE通路的调控作用。体外（in vitro）实验中，我们通过蛋白质印迹（Western blot，WB）与酶联免疫吸附实验（enzyme-linked immunosorbent assay，ELISA）定量检测了T细胞内炎症标志物（肿瘤坏死因子-α（TNF-α）、白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6））的表达变化；体内（in vivo）实验则采用免疫组织化学与流式细胞术，评估了上述细胞因子的表达水平及T细胞计数的改变情况。生物信息学分析结果显示，STIM1在TN患者体内呈现显著过表达，凸显了其在TN病因发生与疾病进展中的核心地位。体外与体内实验结果均证实了STIM1对SOCE通路的调控功能。进一步研究表明，STIM1可介导T淋巴细胞中SOCE诱导的炎症细胞因子释放，这一过程是TN发生发展的关键环节。组织学、超微结构及行为学分析提供的佐证进一步强化了STIM1介导的SOCE与T淋巴细胞驱动的炎症反应在TN发病机制中的关联。本研究提供了全新证据，表明STIM1是T淋巴细胞中SOCE与炎症细胞因子释放的关键调控因子，在三叉神经痛的发病机制中发挥重要作用。本研究结果不仅加深了人们对TN分子基础的理解，同时也为靶向治疗策略的开发开辟了新的方向。