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新疆哈萨克族原发性高血压患者外周血淋巴细胞微小RNA表达谱的生物信息学分析

王彦^1,2, 张亮^1,2,3, 杨瑞^1,2,3, 单莉娅^1,2,3, 王璐^1,2, 张莹莹^1,2, 李新芝^1,2,4, 马克涛^1,2,3,*

¹石河子大学医学院新疆地方与民族高发病教育部重点实验室，石河子 832000；²石河子大学医学院国家卫健委中亚高发病防治重点实验室，石河子 832000；³石河子大学医学院生理学教研室，石河子 832000；⁴石河子大学医学院病理生理学教研室，石河子 832000

摘要

本研究旨在探讨新疆哈萨克族原发性高血压患者和健康者外周血淋巴细胞中微小RNA (microRNA, miRNA)差异表达谱，为高血压的发生机制研究提供理论基础。将2016年4月至2019年5月新疆石河子大学医学院第一附属医院心内科住院部及门诊就诊的30例哈萨克族原发性高血压患者作为高血压组，将30例哈萨克族健康者作为对照组。比较6个哈萨克族高血压患者与6个配对的正常血压者外周血淋巴细胞中miRNA表达谱，对差异表达的miRNA进行聚类、GSEA富集、靶基因预测和靶基因注释等生物信息学分析，并进行qRT-PCR验证。结果显示，与对照组相比，高血压组筛选出73个差异表达miRNA，其中39个miRNA表达上调，34个miRNA表达下调；通过GSEA富集分析共筛选出11个与高血压相关的miRNA，分别是hsa-miR-100-5p、hsa-miR-150-5p、hsa-miR-299-5p、hsa-miR-299-3p、hsa-miR-296-5p、hsa-miR-196b-5p、hsa-miR-503-5p、hsa-miR-628-5p、hsa-miR-874-3p、hsa-miR-543、hsa-miR-940。通过qRT-PCR检验发现，与对照组相比，高血压组hsa-miR-100-5p、hsa-miR-299-5p、hsa-miR-299-3p、hsa-miR-196b-5p、hsa-miR-503-5p、hsa-miR-628-5p、hsa-miR-543表达上调，hsa-miR-150-5p、hsa-miR-296-5p、hsa-miR-874-3p、hsa-miR-940表达下调。基因芯片中的表达趋势和qRT-PCR验证结果一致。利用在线数据库对11个与高血压相关的miRNA进行预测，共预测出1 647个靶基因；GO功能富集分析显示，在生物过程方面，高血压相关靶基因主要与神经系统发育、细胞定位、细胞代谢过程的调节、神经元的产生以及生物过程的正调控等方面相关；在细胞组成方面，主要与膜界细胞器、细胞质、细胞内膜结合的细胞器、突触部分、神经元部分、核质等相关；在分子功能方面，主要与蛋白质结合、转录调控区DNA结合、RNA聚合酶II调节区DNA结合、转录调节活性、离子结合等方面相关；KEGG富集分析显示p53信号通路、心肌细胞中的肾上腺素能信号、cAMP信号通路、内分泌和其他因子调节的钙重吸收、mTOR信号通路、醛固酮调节的钠重吸收、TGF-β信号通路可能与高血压的发生和发展相关。以上结果提示，哈萨克族原发性高血压患者与健康者外周血淋巴细胞中miRNA表达存在差异，筛选出的miRNA可能与哈萨克族原发性高血压的发生和发展相关，其在高血压中的作用机制有待进一步探索验证。

关键词： 高血压; 哈萨克族; 淋巴细胞; microRNA; 表达谱; 生物信息学分析

Bioinformatics analysis of the microRNA expression profile in the peripheral blood lymphocytes of Kazakh patients with essential hypertension in Xinjiang

WANG Yan^1,2, ZHANG Liang^1,2,3, YANG Rui^1,2,3, SHAN Li-Ya^1,2,3, WANG Lu^1,2, ZHANG Ying-Ying^1,2, LI Xin-Zhi^1,2,4, MA Ke-Tao^1,2,3,*

¹Key Laboratory of Xinjiang Endemic and Ethnic Diseases, Ministry of Education, Shihezi University School of Medicine, Shihezi 832000, China；²NHC Key Laboratory of Prevention and Treatment of Central Asia High Incidence Diseases, Shihezi University School of Medicine, Shihezi 832000, China；³Department of Physiology, Shihezi University School of Medicine, Shihezi 832000, China；⁴Department of Pathophysiology, Shihezi University School of Medicine, Shihezi 832000, China

Abstract

This study aimed to investigate the differential expression profiles of microRNAs (miRNAs) in peripheral blood lymphocytes between patients with essential hypertension and healthy individuals in Xinjiang Kazakh and to provide insight into the mechanism involved in the pathogenesis of hypertension in this ethnic group. From April 2016 to May 2019, 30 Kazakh patients with essential hypertension in the inpatient and outpatient departments of Cardiology, First Affiliated Hospital of Shihezi University were used as the hypertension group; 30 healthy Kazakh patients were used as the control group. The miRNA expression profiles in peripheral blood lymphocytes of 6 Kazakh hypertensive patients and 6 matched healthy individuals were compared, and the differentially expressed miRNAs were analyzed by cluster analysis, GSEA enrichment analysis, target gene prediction, target gene annotation and other bioinformatics analyses. In addition, qRT-PCR was used to verify the differentially expressed miRNAs. The results showed that compared with the control group, 73 differentially expressed miRNAs were identified in the hypertension group, of which 39 miRNAs were up-regulated and 34 miRNAs were down-regulated. A total of 11 miRNAs related to hypertension were screened by GSEA enrichment analysis, including hsa-miR-100-5p, hsa-miR-150-5p, hsa-miR-299-5p, hsa-miR-299-3p, hsa-miR-296-5p, hsa-miR-196b-5p, hsa-miR-503-5p, hsa-miR-628-5p, hsa-miR-874-3p, hsa-miR-543 and hsa-miR-940. qRT-PCR test found that the expression of hsa-miR-100-5p, hsa-miR-299-5p, hsa-miR-299-3p, hsa-miR-196b-5p, hsa-miR-503-5p, hsa-miR-628-5p and hsa-miR-543 was up-regulated, while the expression of hsa-miR-150-5p, hsa-miR-296-5p, hsa-miR-874-3p and hsa-miR-940 was down-regulated in the hypertension group compared with the control group. The expression trend in the gene chip was consistent with the results verified by qRT-PCR. Using online database to predict target genes of 11 miRNAs related to hypertension, we found that a total of 1 647 target genes might be regulated by these 11 miRNAs. GO function enrichment showed that (a) in biological processes, the predicted hypertension related target genes are mainly relevant to nervous system development, cellular localization, regulation of cellular metabolic process, generation of neurons and positive regulation of biological process; (b) In terms of cellular components, they are mainly related to membrane-bounded organelle, cytoplasm, intracellular membrane-bounded organelle, synapse part, neuron part, and nucleoplasm; (c) In terms of molecular function, they are mainly related to protein binding, transcription regulatory region DNA binding, RNA polymerase II regulatory region DNA binding, transcription regulator activity, and ion binding. KEGG enrichment analysis showed that the p53 signaling pathway, adrenergic signaling in cardiomyocytes, cAMP signaling pathway, TGF-β signaling pathway, endocrine and other factor-regulated calcium reabsorption, mTOR signaling pathway, and aldosterone-regulated sodium reabsorption may be related to the occurrence and development of hypertension. In conclusion, there are significant differences in the expression of miRNAs in peripheral blood lymphocytes between Kazakh patients with essential hypertension and healthy people. The differentially expressed miRNAs may be related to the occurrence and development of essential hypertension in Kazakh. However, the underlying mechanism needs to be further explored and verified.

Key words: Hypertension; Kazakh; lymphocytes; microRNA; expression profile; bioinformatics analysis

收稿日期：　　录用日期：

通讯作者：马克涛　　E-mail: maketao@hotmail.com

DOI: 10.13294/j.aps.2022.0077

引用本文：

王彦, 张亮, 杨瑞, 单莉娅, 王璐, 张莹莹, 李新芝, 马克涛. 新疆哈萨克族原发性高血压患者外周血淋巴细胞微小RNA表达谱的生物信息学分析[J]. 生理学报 2022; 74 (5): 751-762.

WANG Yan, ZHANG Liang, YANG Rui, SHAN Li-Ya, WANG Lu, ZHANG Ying-Ying, LI Xin-Zhi, MA Ke-Tao. Bioinformatics analysis of the microRNA expression profile in the peripheral blood lymphocytes of Kazakh patients with essential hypertension in Xinjiang. Acta Physiol Sin 2022; 74 (5): 751-762 (in Chinese with English abstract).