The Critical Biomarkers Identification of Insulin Signaling Involved in Initiating cAMP Signaling Mediated Salivary Secretion in Sjogren Syndrome: Transcriptome Sequencing in NOD Mice Model.

IF 3.7 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS Biological Procedures Online Pub Date : 2022-12-27 DOI:10.1186/s12575-022-00189-5

Bo Chen, Jiannan Zhou, Tianjiao Mao, Tingting Cao, Shilin Hu, Wenqi Zhang, Xueyang Li, Xiuni Qin, Xintong Liu, Nobumoto Watanabe, Jiang Li

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Abstract

Background: Sjogren's syndrome (SS) is an autoimmune disorder characterized by the destruction of exocrine glands, resulting in dry mouth and eyes. Currently, there is no effective treatment for SS, and the mechanisms associated with inadequate salivary secretion are poorly understood.

Methods: In this study, we used NOD mice model to monitor changes in mice's salivary secretion and water consumption. Tissue morphology of the submandibular glands was examined by H&E staining, and Immunohistochemical detected the expression of AQP5 (an essential protein in salivary secretion). Global gene expression profiling was performed on submandibular gland tissue of extracted NOD mice model using RNA-seq. Subsequently, a series of bioinformatics analyses of transcriptome sequencing was performed, including differentially expressed genes (DEGs) identification, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, PPI network construction, hub gene identification, and the validity of diagnostic indicators using the dataset GSE40611. Finally, IFN-γ was used to treat the cells, the submandibular gland tissue of NOD mice model was extracted, and RT-qPCR was applied to verify the expression of hub genes.

Results: We found that NOD mice model had reduced salivary secretion and increased water consumption. H&E staining suggests acinar destruction and basement membrane changes in glandular tissue. Immunohistochemistry detects a decrease in AQP5 immunostaining within acinar. In transcriptome sequencing, 42 overlapping DEGs were identified, and hub genes (REN, A2M, SNCA, KLK3, TTR, and AZGP1) were identified as initiating targets for insulin signaling. In addition, insulin signaling and cAMP signaling are potential pathways for regulating salivary secretion and constructing a regulatory relationship between target-cAMP signaling-salivary secretion.

Conclusion: The new potential targets and signal axes for regulating salivary secretion provide a strategy for SS therapy in a clinical setting.

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干燥综合征中参与启动cAMP信号介导的唾液分泌的胰岛素信号的关键生物标志物鉴定:NOD小鼠模型的转录组测序。

背景:干燥综合征(SS)是一种自身免疫性疾病，其特征是外分泌腺的破坏，导致口干和眼睛干燥。目前，对SS没有有效的治疗方法，与唾液分泌不足相关的机制也知之甚少。方法:本研究采用NOD小鼠模型，监测小鼠唾液分泌和饮水量的变化。H&E染色检测颌下腺组织形态，免疫组化检测唾液分泌必需蛋白AQP5的表达。采用RNA-seq对提取的NOD小鼠颌下腺组织进行全局基因表达谱分析。随后，对转录组测序进行了一系列生物信息学分析，包括差异表达基因(DEGs)鉴定、基因本体(GO)和京都基因与基因组百科全书(KEGG)富集分析、PPI网络构建、枢纽基因鉴定以及使用数据集GSE40611诊断指标的有效性。最后用IFN-γ处理细胞，提取NOD小鼠颌下腺模型组织，采用RT-qPCR验证hub基因的表达。结果:我们发现NOD小鼠模型唾液分泌减少，饮水量增加。H&E染色提示腺泡破坏，基底膜改变。免疫组化检测到腺泡内AQP5免疫染色降低。在转录组测序中，鉴定出42个重叠的deg，中心基因(REN, A2M, SNCA, KLK3, TTR和AZGP1)被鉴定为胰岛素信号传导的起始靶点。此外，胰岛素信号和cAMP信号是调节唾液分泌的潜在途径，构建了靶点-cAMP信号-唾液分泌的调控关系。结论:涎腺分泌调节的新靶点和信号轴为临床治疗SS提供了新的策略。

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来源期刊

Biological Procedures Online 生物-生化研究方法

CiteScore

10.50

自引率

0.00%

发文量

审稿时长

>12 weeks

期刊介绍： iological Procedures Online publishes articles that improve access to techniques and methods in the medical and biological sciences. We are also interested in short but important research discoveries, such as new animal disease models. Topics of interest include, but are not limited to: Reports of new research techniques and applications of existing techniques Technical analyses of research techniques and published reports Validity analyses of research methods and approaches to judging the validity of research reports Application of common research methods Reviews of existing techniques Novel/important product information Biological Procedures Online places emphasis on multidisciplinary approaches that integrate methodologies from medicine, biology, chemistry, imaging, engineering, bioinformatics, computer science, and systems analysis.