生物信息学分析哮喘病人气道上皮microRNA的表达

生物信息学分析哮喘病人气道上皮microRNA的表达

杨静

电子科技大学医学院附属妇女儿童医院•成都市妇女儿童中心医院 四川省成都市  610073

关键词

哮喘，microRNA，基因芯片

摘要

目的：本研究旨在通过生物信息学研究初步筛选GEO数据库中哮喘病人气道上皮中增高和降低的microRNA。

方法：通过查询GEO数据库中健康人和哮喘病人气道上皮的microRNA表达的基因微阵列芯片数据（GSE25230和GSE142237），分析芯片中增高大于0.5倍(FC>1.5)和降低0.25倍（FC<0.75）的microRNA数据,分别将两个芯片中增高的microRNA和降低的microRNA数据做韦恩图取交集，以筛选出2个芯片中同时增高和降低的microRNA。

结果：在2个芯片中，与健康对照组相比，哮喘组气道上皮细胞中有15个microRNA增高(miR-100-5p, miR-30a-5p, miR-200a-3p,miR-27b-5p, let-7f-5p, miR-30d-5p ,miR-335-5p, miR-1293, miR-21-5p, miR-345-5p, miR-1244, miR-99a-5p, miR-130b-5p, miR-181a-3p, miR-374a-3p)。有21个microRNA下降（miR-193b ,miR-877, miR-485-3p, miR-720, miR-1236, miR-1291, miR-675-5p, miR-1207-5p, miR-519e-5p, miR-941, miR-450a-5p, miR-630, miR-185-3p, miR-522-5p, miR-936, miR-149-3p, miR-124-5p, miR-498, miR-638 ,miR-885, miR-105-5p）。

结论：我们通过分析2个基因芯片，获得了15个同时增高和21个同时降低的microRNA。

Abstract

Objective: This study is aimed at finding out increased and decreased microRNAs in bronchial epithelium of asthmatic patients by analyzing GEO datasets with bioinformatics.

Method: In GEO datasets, chips analyzing microRNAs in bronchial epithelium of asthmatics were screened, and we then found the two chips, GSE25230 and GSE142237. microRNAs increased by 0.5 fold change or decreased by 0.25 fold change were analyzed and listed in the two chips. To get commen microRNAs in the two chips, the increased and decreased microRNAs were analyzed with Venn graph separately.

Result: In the two chips, 15 microRNAs were increased in bronchial epithelium of asthmatics compared with healthy controls (miR-100-5p, miR-30a-5p, miR-200a-3p,miR-27b-5p, let-7f-5p, miR-30d-5p ,miR-335-5p, miR-1293, miR-21-5p, miR-345-5p, miR-1244, miR-99a-5p, miR-130b-5p, miR-181a-3p, miR-374a-3p).;21 microRNAs were decreased (miR-193b ,miR-877, miR-485-3p, miR-720, miR-1236, miR-1291, miR-675-5p, miR-1207-5p, miR-519e-5p, miR-941, miR-450a-5p, miR-630, miR-185-3p, miR-522-5p, miR-936, miR-149-3p, miR-124-5p, miR-498, miR-638 ,miR-885, miR-105-5p).

Conclusion: In the two chips, 15 microRNAs were increased and 21 microRNAs were decreased at the same time.

前言

哮喘作为最流行的非传染性疾病之一，影响着全世界大约有3亿人的生活，每年造成许多患者死亡，患病人群分布在各个年龄、种族和民族。作为一种常见的慢性气道炎症性疾病，长期的治疗费用昂贵，尤其是给低收入人群造成了巨大的经济负担 [1,2]。近年来，哮喘的患病人数在全世界范围内都是呈增加的状态，尤其是西方发达国家[3,4]。哮喘的症状主要包括：气喘、呼吸困难、胸闷和咳嗽，主要是由气管中的气流阻塞引起。气道组织中粘液的过度分泌是哮喘和慢性阻塞性肺病(COPD)的主要病理特征。近年来，关于哮喘的发病机制研究颇丰，其中microRNA在哮喘的发病机制中发挥着重要作用[5,6]。microRNA是一类长度在20-22个核苷酸的非编码RNA,在转录后水平通过序列特异性结合mRNA,抑制其翻译或导致其降解，从而调控基因的表达[7]。本研究旨在筛选GEO数据库中研究哮喘患者气道上皮细胞中microRNA表达的芯片，为后续研究奠定基础。

1资料与方法

1.1资料

查询基因芯片数据库GEO Datasets(https://www.ncbi.nlm.nih.gov/)，搜索与哮喘和microRNA的芯片，从中筛选出研究健康对照和哮喘患者气道上皮细胞microRNA的芯片（GSE25230和GSE142237），下载数据备用。

1.2方法

首先使用EXCEL处理数据并排序，再使用软件Bioinformatics&EvolutionaryGenomics（http://bioinformatics.psb.ugent.be/webtools/Venn/）对两组芯片中FC>1.5和FC<0.75 的microRNA分别取交集；通过microRNA查询网站miRBase（https://www.mirbase.org/）查询microRNA的序列，分析取交集的microRNA数据是否一致，最终我们准确得到了2个芯片中增高大于0.5倍(FC>1.5)和降低0.25倍（FC<0.75）的microRNA数据。

2结果

（1）在GSE25230和GSE142237芯片中，与健康组相比，哮喘组气道上皮细胞中有15个microRNA增高，且FC>1.5倍：miR-100-5p, miR-30a-5p, miR-200a-3p,miR-27b-5p, let-7f-5p, miR-30d-5p ,miR-335-5p, miR-1293, miR-21-5p, miR-345-5p, miR-1244, miR-99a-5p, miR-130b-5p, miR-181a-3p, miR-374a-3p。（图1）

（2）在GSE25230和GSE142237芯片中，与健康组相比，哮喘组气道上皮细胞中有21个microRNA降低，且FC<0.75倍：miR-193b ,miR-877, miR-485-3p, miR-720, miR-1236, miR-1291, miR-675-5p, miR-1207-5p, miR-519e-5p, miR-941, miR-450a-5p, miR-630, miR-185-3p, miR-522-5p, miR-936, miR-149-3p, miR-124-5p, miR-498, miR-638 ,miR-885, miR-105-5p。（图2）

3 讨论

我们通过查找GEO数据库中与哮喘相关的microRNA芯片，筛选出检测健康对照组和哮喘患者组气道上皮细胞microRNA表达的芯片（GSE25230和GSE142237），并分析其中增高0.5倍和降低0.25倍的microRNA，分别对两个芯片中增高和降低的microRNA取交集，扩大了样本数，提高了筛选出的microRNA的准确度。筛选出的这些microRNA可以为后续做哮喘的病理机制研究提供思路。

参考文献

[1] Masoli, M., et al., The global burden of asthma: executive summary of the GINA Dissemination Committee report. Allergy, 2004. 59(5): p. 469-78.

[2] Pearce, N., et al., Worldwide trends in the prevalence of asthma symptoms: phase III of the International Study of Asthma and Allergies in Childhood (ISAAC). Thorax, 2007. 62(9): p. 758-66.

[3] Foronda, C.L., et al., Healthcare providers' experiences with gaps, barriers, and facilitators faced by family caregivers of children with respiratory diseases. J Pediatr Nurs, 2020. 52: p. 49-53.

[4] Ferkol, T. and D. Schraufnagel, The global burden of respiratory disease. Ann Am Thorac Soc, 2014. 11(3): p. 404-6.

[5] Cañas JA, Rodrigo-Muñoz JM, Gil-Martínez M, Sastre B, del Pozo V. Exosomes: a key piece in asthmatic infammation. Int J Mol Sci. 2021;22(2):963.

[6] Bélanger É, Madore A-M, Boucher-Lafeur A-M, Simon M-M, Kwan T, Pastinen T, et al. Eosinophil microRNAs play a regulatory role in allergic diseases included in the atopic march. Int J Mol Sci. 2020;21(23):9011.

[7] Kim VN, Han J, Siomi MC. Biogenesis of small RNAs in animals. Nat Rev Mol Cell Biol. 2009;10(2):126–39.