基于单细胞测序的草酸钙肾结石大鼠髓袢细胞亚群基因表达谱特征分析

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摘要目的：探讨草酸钙肾结石大鼠模型肾脏组织髓袢细胞亚群基因表达谱特征。方法：应用乙二醇氯化铵诱导法构建草酸钙肾结石大鼠模型，通过Von Kossa′s组织染色验证模型构建成功。单细胞测序方法鉴定髓袢细胞，对比分析草酸钙肾结石大鼠模型肾脏组织髓袢细胞亚群基因表达谱特征，应用生物信息学方法进行差异表达基因的GO、KEGG以及GSEA富集分析。结果：相较于对照组，实验组大鼠肾脏组织中发现钙盐沉积区域呈黑色或棕黑色，表明草酸钙肾结石大鼠模型构建成功。单细胞测序分析发现，在实验组髓袢细胞亚群共有差异表达基因3 510个，其中880个表达上调，2 630个表达下调；其中LOC499584，白介素7（interleukin 7,Il7），双特异性磷酸酶1（dual specificity phosphatase 1,Dusp1）以及基质Gla蛋白（matrix Gla protein，Mgp）表达上调最为显著；HORMA结构域蛋白质2（HORMA domain containing 2，Hormad2），LOC361914，JUN二聚体蛋白2（Jun dimerization protein 2，Jdp2）以及生长抑素-B和血小板反应蛋白1型结构域蛋白（somatomedin-B and thrombospondin type-1 domain-containing protein，Sbspon）表达下调最为显著。KEGG分析发现实验组髓袢细胞亚群氧化磷酸化、代谢途径、丙酸代谢、自噬、溶酶体以及胞吞作用等信号通路发生显著改变。结论：本研究首次揭示了草酸钙肾结石大鼠模型肾脏组织髓袢细胞亚群基因表达谱特征，为肾结石的形成及其介导的肾损伤研究提供了一定的参考依据。

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关键词 ： , 肾结石 83, 179), font-family: ????, font-weight: 700, text-align: -webkit-right, background-color: rgb(212, 226, 255), ')" href="#">">, 单细胞测序 83, 179), font-family: ????, font-weight: 700, text-align: -webkit-right, background-color: rgb(212, 226, 255), ')" href="#">">, 表达谱 83, 179), font-family: ????, font-weight: 700, text-align: -webkit-right, background-color: rgb(212, 226, 255), ')" href="#">">, 髓袢 83, 179), font-family: ????, font-weight: 700, text-align: -webkit-right, background-color: rgb(212, 226, 255), ')" href="#">">, 生物信息学

Abstract： Objective： To explore the gene expression profiles of Henle-loop cell subset in renal tissue of the calcium oxalate kidney stone rat model. Methods： The rat model of calcium oxalate kidney stone was constructed by ethylene glycol and 1% ammonium chloride. The model was validated by Von Kossa′s staining. Henle-loop cells were identified using single-cell sequencing method. The gene expression profiles of Henle-loop cell subsets in renal tissue of calcium oxalate kidney stone rat model were performed via comparative analysis. The enrichment analysis of differential genes was performed through GO, KEGG, and GSEA bioinformatics tools. Results： Compared with the control group, black or brownish black staining of calcium salt deposition were observed in the kidney tissue of the experimental group rats, indicating the successful construction of the calcium oxalate kidney stone rat model. Single cell sequencing analysis revealed that there was a total of 3 510 differentially expressed genes in the Henle-loop cell subset of the experimental group, of which 880 were upregulated and 2 630 were downregulated. LOC499584, Il7, dual specificity phosphatase 1 (Dusp1), and matrix Gla protein (Mgp) were upregulated most significantly, while HORMA domain containing 2 (Hormad2), LOC361914, Jun dimerization protein 2 (Jdp2), and somatomedinB and thrombospondin type-1 domain-containing protein (Sbspon) were downregulated most significantly. KEGG analysis revealed significant changes in signaling pathways such as oxidative phosphorylation, metabolism pathways, propanoate metabolism, autophagy, lysosome and endocytosis in the experimental group of Henle-loop cells. Conclusion： This study reveals for the first time the gene expression profile characteristics of Henleloop cell subsets in renal tissue of the calcium oxalate kidney stone rat model, providing a molecular basis for the investigation of kidney stone formation and related kidney injury.

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收稿日期: 2023-12-18

基金资助: 广东省基础与应用基础研究基金(省企联合，2022A1515220106）；广东省医学科学技术研究基金项目（A2023281）

通讯作者: 王铸（通讯作者），副研究员，E-mail: wangzhu_1223@163.com

作者简介: 许婷（1983—），女，主管护师，主要从事泌尿系结石疾病研究

引用本文:

许婷1，张颖1，邓琼1,2，梁辉1，王铸1,2. 基于单细胞测序的草酸钙肾结石大鼠髓袢细胞亚群基因表达谱特征分析[J]. 江苏大学学报:医学版, 2024, 34(02): 104-110. XU Ting1, ZHANG Ying1, DENG Qiong1,2, LIANG Hui1, WANG Zhu1,2. The gene expression profiles of Henle-loop cell subset in renal tissue of the calcium oxalate kidney stone rat model based on single-cell sequencing analysis. Journal of Jiangsu University(Medicine Edition), 2024, 34(02): 104-110.

链接本文:

http://zzs.ujs.edu.cn/xbyxb/CN/ 或 http://zzs.ujs.edu.cn/xbyxb/CN/Y2024/V34/I02/104

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