Xie Dong, Liu Yang, Xu Fang-Biao, Zhang Jin-Sheng
The Third Clinical Medical College, Henan University of Chinese Medicine, Zhengzhou, China.
Department of Encephalopathy, The Third Affiliated Hospital of Henan University of Chinese Medicine, Zhengzhou, China.
Front Neurol. 2025 Jan 27;15:1513357. doi: 10.3389/fneur.2024.1513357. eCollection 2024.
The escalating incidence of ischemic stroke (IS) exerts a heavy toll on global health. Aging, a prominent risk factor, implicates the senescence-associated secretory phenotype (SASP) in IS pathogenesis. We postulated that alterations in SASP-related factor expression during IS correlate with remodeling of intercellular interaction networks and disease advancement. The present study endeavored to preliminarily dissect the SASP-IS nexus via combined bulk and single-cell transcriptome analysis.
Aggregated expression profiles from human peripheral blood bulk chips and MCAO mouse single-cell sequencing data, followed by SASP gene analysis. Executed protein interaction network and enrichment assays. Investigated immune infiltration in stroke patients, managed quality control and annotation of single-cell data, cherry-picked central cells based on SASP scores, unearthed essential genes via enrichment analysis, conducted pseudo-time and intercellular communication studies, and prognosticated drugs for hub genes. Finally, authenticated core gene expression in serum of MCAO and Sham rats using real-time fluorescent polymerase chain reaction (RT-qPCR).
Fourteen hub genes were discerned. Seven cell types were annotated in MCAO mouse peripheral blood single-cell data. Basophils exhibited the highest SASP scores, with Lcp1 upregulated and Ccl3 downregulated in basophils of the MCAO group. Enrichment analysis divulged a significant association of Ccl3 with the cell apoptosis pathway and Lcp1 with immune responses. The Ccl3 gene is pivotal in basophils and basophil-neutrophil crosstalk. Additionally, we forecasted nagrestipen's regulatory function on Ccl3. RT-qPCR demonstrated a marked elevation in Lcp1 mRNA and a pronounced reduction in Ccl3 in the MCAO group relative to the Sham group.
The Ccl3 gene in basophils and its immune cell interaction is a linchpin in the IS immune microenvironment. Ccl3 and Lcp1 might potentially modulate IS progression by influencing SASP, proffering novel prospects for IS clinical diagnosis and treatment.
缺血性中风（IS）发病率不断上升，给全球健康带来沉重负担。衰老作为一个突出的风险因素，在IS发病机制中涉及衰老相关分泌表型（SASP）。我们推测，IS期间SASP相关因子表达的改变与细胞间相互作用网络的重塑和疾病进展相关。本研究试图通过批量和单细胞转录组分析相结合的方法初步剖析SASP与IS的关系。
整合人类外周血批量芯片的表达谱和MCAO小鼠单细胞测序数据，随后进行SASP基因分析。执行蛋白质相互作用网络和富集分析。研究中风患者的免疫浸润情况，对单细胞数据进行质量控制和注释，根据SASP评分挑选核心细胞，通过富集分析挖掘关键基因，进行伪时间和细胞间通讯研究，并预测枢纽基因的药物。最后，使用实时荧光聚合酶链反应（RT-qPCR）验证MCAO和假手术大鼠血清中的核心基因表达。
识别出14个枢纽基因。在MCAO小鼠外周血单细胞数据中注释了7种细胞类型。嗜碱性粒细胞的SASP评分最高，MCAO组嗜碱性粒细胞中Lcp1上调而Ccl3下调。富集分析揭示Ccl3与细胞凋亡途径显著相关，Lcp1与免疫反应显著相关。Ccl3基因在嗜碱性粒细胞和嗜碱性粒细胞-中性粒细胞相互作用中起关键作用。此外，我们预测了那格列净对Ccl3的调节作用。RT-qPCR结果显示，与假手术组相比，MCAO组Lcp1 mRNA显著升高，Ccl3显著降低。
嗜碱性粒细胞中的Ccl3基因及其免疫细胞相互作用是IS免疫微环境中的关键因素。Ccl3和Lcp1可能通过影响SASP来调节IS进展，为IS的临床诊断和治疗提供了新的前景。
