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白色念珠菌性眼内炎小鼠模型中外泌体的蛋白质组学分析。

Proteomic profiling of exosomes in a mouse model of Candida albicans endophthalmitis.

机构信息

Jhaveri Microbiology Centre, Brien Holden Eye Research Centre, L. V. Prasad Eye Institute, Hyderabad, Telangana, India; Manipal Academy of Higher Education, Karnataka, India.

Department of Biomedical Engineering, Indian Institute of Technology Hyderabad, Telangana, India.

出版信息

Exp Cell Res. 2022 Aug 15;417(2):113222. doi: 10.1016/j.yexcr.2022.113222. Epub 2022 May 24.

Abstract

Exosomes play pivotal roles in intercellular communication, and pathophysiological functions. In this study, we aimed to understand the role of exosomal proteome derived from C. albicans infected mice (C57BL/6) eyeball. Exosomes were characterized by Dynamic Light Scattering and Western blot, quantified and subjected to LC-MS/MS and cytokine quantification by ELISA. The average size of exosomes was 170-200 nm with number of exosomes amounted to 1.42 × 10 in infected set compared to control (1.24 × 10). Western blot was positive for CD9, CD63 and CD81 confirming the presence of exosomes. IL-6, IL1β, TNF-α, and IFN-γ levels were significantly elevated in infected eye at 72 h.p.i. Proteomic analysis identified 42 differentially expressed proteins, of these 37 were upregulated and 5 were downregulated. Gene Ontology (GO) revealed enrichment of cell adhesion, cytoskeleton organisation, and cellular response proteins such as aquaporin-5, gasdermin-A, CD5 antigen-like, Catenin, V-ATPase, and vesicle associated protein. Additionally, KEGG pathway analysis indicated the association of metabolic and carbon signalling pathways with exosomes from C. albicans infected eye. The protein cargo in exosomes released during endophthalmitis with C. albicans seems to play a unique role in the pathogenesis of the disease and further validations with larger cohort of patients is required to confirm them as biomarkers.

摘要

外泌体在细胞间通讯和病理生理功能中发挥着关键作用。在本研究中,我们旨在了解白色念珠菌感染小鼠(C57BL/6)眼球来源的外泌体蛋白质组的作用。通过动态光散射和 Western blot 对外泌体进行了特征分析,通过 LC-MS/MS 和 ELISA 对细胞因子进行了定量和分析。与对照组(1.24×10)相比,感染组中 170-200nm 大小的外泌体平均数量为 1.42×10,数量增加。Western blot 检测到 CD9、CD63 和 CD81 呈阳性,证实了外泌体的存在。在感染后 72 小时,眼内的 IL-6、IL1β、TNF-α 和 IFN-γ 水平显著升高。蛋白质组学分析鉴定出 42 种差异表达蛋白,其中 37 种上调,5 种下调。基因本体论(GO)揭示了细胞粘附、细胞骨架组织和细胞反应蛋白的富集,如水通道蛋白-5、gasdermin-A、CD5 抗原样、连环蛋白、V-ATPase 和囊泡相关蛋白。此外,KEGG 通路分析表明,与白色念珠菌感染眼外泌体相关的代谢和碳信号通路的关联。在白色念珠菌性眼内炎中释放的外泌体中的蛋白质货物似乎在疾病的发病机制中发挥着独特的作用,需要对更大的患者队列进行进一步验证,以确认它们作为生物标志物。

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