种植体周围炎中铁死亡的生物信息学分析与实验验证

doi:10.13591/j.cnki.kqyx.2024.07.008

摘要/Abstract

摘要：

目的探究种植体周围炎中与铁死亡相关的关键基因,探究调控种植体周围炎的潜在机制。方法从GEO数据库中获取多个芯片数据,筛选出差异表达基因,进行GO及KEGG分析,STRING网站构建差异基因PPI网络;利用测试集对关键基因进行验证,并判断关键基因的诊断价值。通过免疫浸润分析获得22种免疫细胞在种植体周围炎组织中的含量及比例。对关键基因进行qRT-PCR与Western Blot(WB)验证。结果种植体周围炎组织与正常牙龈组织共1 138个差异基因,其中29个为铁死亡相关差异基因;种植体周围炎组织的基因表达主要涉及免疫反应激活等过程;铁死亡相关差异基因中的5个关键基因,即SOX2、GJA1、IL1B、GPX2和CHAC1,在种植体周围炎组织中差异表达并有较高诊断价值。免疫浸润分析结果显示,在种植体周围炎组织中记忆B细胞、浆细胞等免疫细胞显著变化。qRT-PCR实验与WB实验证实关键基因mRNA与其转录的蛋白有明显的差异表达。结论利用生物信息学分析与生物学验证筛选出种植体周围炎和铁死亡间的差异基因,为种植体周围炎的研究提供新思路。

关键词: GEO数据库, 生物信息学, 种植体周围炎, 铁死亡, 差异基因

Abstract:

Objective To investigate the key genes associated with ferroptosis in peri-implantitis and explore the potential mechanisms regulating peri-implantitis. Methods Several datasets were obtained from the GEO database. Differential expressed genes were screened, and GO and KEGG analyses were performed. A PPI network was constructed using the STRING website. Key genes were validated using a test set, and the diagnostic value of key genes was determined. The content and proportion of 22 immune cells in peri-implantitis tissues were obtained through immune infiltration analysis. Key genes were validated by qRT-PCR and Western Blot(WB). Results There were 1 138 differential genes between peri-implantitis tissues and normal gingival tissues, of which 29 were related to ferroptosis. The gene expression in peri-implantitis tissues mainly involved processes such as immune response activation. Five key genes in the ferroptosis-related differential genes, namely SOX2, GJA1, IL1B, GPX2 and CHAC1, were differentially expressed in peri-implantitis tissues and had high diagnostic value. Immune infiltration analysis showed significant changes in immune cells such as memory B cells and plasma cells in peri-implantitis tissues. qRT-PCR and WB confirmed significant differential expression of mRNA and the protein transcribed by key genes. Conclusion Differential genes between peri-implantitis and ferroptosis are screened using bioinformatics analysis and biological validation, providing new insights into the study on peri-implantitis.

Key words: GEO database, bioinformatics, peri-implantitis, ferroptosis, differential expressed genes

中图分类号:

R318.04

张哲维, 汪姣宏, 吴维, 董硕, 李国情, 汤春波. 种植体周围炎中铁死亡的生物信息学分析与实验验证[J]. 口腔医学, 2024, 44(7): 527-535.

ZHANG Zhewei, WANG Jiaohong, WU Wei, DONG Shuo, LI Guoqing, TANG Chunbo. Bioinformatics analysis and experimental validation of ferroptosis in peri-implantitis[J]. Stomatology, 2024, 44(7): 527-535.

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基因名称	引物序列(5'—3')
SOX2-F	GACAGTTACGCGCACATGAA
SOX2-R	GTTCATGTAGGTCTGCGAGC
GJA1-L	CTCGCCTATGTCTCCTCCTG
GJA1-R	CCCAGTTTTGCTCACTTGCT
GPX2-L	ATGTGAGGTGAATGGGCAGA
GPX2-R	ATGATGAGCTTGGGATCGGT
CHAC1-L	GATCATGAGGGCTGCACTTG
CHAC1-R	CCACAGAGCTGCATGAAGTC
IL1B-L	CGAATCTCCGACCACCACTA
IL1B-R	AGCCTCGTTATCCCATGTG

通路	基因数目	富集评分	校正后的归一化的富集评分值	P
造血细胞谱系	91	0.66	2.34	1.03×10^-10
细胞因子与细胞因子受体相互作用	267	0.54	2.19	1.00×10^-10
NF-κB信号通路	96	0.62	2.24	2.61×10^-9
趋化因子信号通路	179	0.52	2.04	7.13×10^-9
类风湿性关节炎	83	0.63	2.22	1.96×10^-8

通路描述	基因数目	富集评分	校正后的归一化的富集评分值	P
B细胞增殖	96	0.72	2.59	1.00×10^-10
B细胞增殖调节	63	0.73	2.45	1.00×10^-10
免疫应答调节细胞表面受体信号通路	291	0.59	2.43	1.00×10^-10
B细胞受体信号通路	64	0.72	2.40	1.00×10^-10
适应性免疫应答	450	0.56	2.40	1.00×10^-10