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转录组测序研究内毒素诱导的人视网膜色素上皮细胞炎症反应的机制
Mechanism of LPS-induced inflammation in human retinal pigment epithelium cells by transcriptome sequencing
- 第三军医大学学报 2019年41卷第3期 页码：192-198
- 作者机构：
1..400016 重庆，重庆医科大学附属第一医院眼科，眼科学重庆市重点实验室Department of Ophthalmology, Chongqing Key Laboratory of Ophthalmology, the First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016
2..401220 重庆，重庆市长寿区人民医院眼科Department of Ophthalmology, People's Hospital of Changshou District, Chongqing, 401220
3..450003 郑州，河南省人民医院，河南省立眼科医院，河南省眼科研究所Henan Provincial Eye Hospital, Henan Eye Institute, Henan Provincial People's Hospital, Zhengzhou, Henan Province, 450003, China
- 作者简介：
- 基金信息：国家自然基金面上项目(81770949);河南省科技攻关项目(182102310145)
DOI：10.16016/j.1000-5404.201809033
中图分类号： R363.2;R392.13;R774.11收稿：2018-09-05，
纸质出版：2019-02-15
- 稿件说明：
移动端阅览
余鹏, 阳琳, 麦雨欣, 等. 转录组测序研究内毒素诱导的人视网膜色素上皮细胞炎症反应的机制[J]. 第三军医大学学报, 2019,41(3):192-198.
Peng YU, Lin YANG, Yuxin MAI, et al. Mechanism of LPS-induced inflammation in human retinal pigment epithelium cells by transcriptome sequencing[J]. Journal of Third Military Medical University, 2019, 41(3): 192-198.
余鹏, 阳琳, 麦雨欣, 等. 转录组测序研究内毒素诱导的人视网膜色素上皮细胞炎症反应的机制[J]. 第三军医大学学报, 2019,41(3):192-198. DOI： 10.16016/j.1000-5404.201809033.
Peng YU, Lin YANG, Yuxin MAI, et al. Mechanism of LPS-induced inflammation in human retinal pigment epithelium cells by transcriptome sequencing[J]. Journal of Third Military Medical University, 2019, 41(3): 192-198. DOI： 10.16016/j.1000-5404.201809033.
摘要
目的
2
转录组测序探讨内毒素(lipopolysaccharide
LPS)诱导的人视网膜色素上皮细胞(retinal pigment epithelium cells
ARPE-19)炎症反应的机制。
方法
2
将ARPE-19细胞分为空白对照组和LPS组
Real-time PCR检测炎症因子IL-6、IL-8和MCP-1的表达
Illumina Hiseq 2500用于转录组测序
DESeq2用于差异表达基因分析
采用GOseq R package和Omicsbean软件分别对差异表达基因进行GO和KEGG富集分析; 再将ARPE-19细胞和hRPE细胞分为空白对照组和LPS组
Real-time PCR验证RNA-seq结果中的关键基因。
结果
2
LPS刺激ARPE-19细胞24 h后
炎症因子IL-6、IL-8和MCP-1在mRNA水平的表达显著升高(
P
<
0.01)。与空白对照组比较
LPS组有2 345个差异表达基因
其中上调的有1 594个
下调的有751个。GO和KEGG富集分析显示LPS激活了免疫相关通路和Toll-样受体、MAPK、HIF-1、TNF、PI3K-Akt、NOD-样受体、mTOR、VEGF信号通路以及转录因子JUN、JUNB、SRF、ATF4和AKT1、SRC基因。
结论
2
LPS诱导的ARPE-19细胞炎症反应可能与PI3K-Akt-mTOR、NOD-样受体、VEGF信号通路及转录因子FOS、JUN、JUNB、SRF和AKT1、SRC基因的激活有关。
Abstract
Objective
2
To investigate the mechanism of lipopolysaccharide (LPS)-induced inflammation in retinal pigment epithelium (ARPE-19) cells by analyzing the transcriptome profiles.
Methods
2
ARPE-19 cells were divided into control group and LPS group. Real-time PCR was performed to detect the expression of inflammatory cytokines IL-6
IL-8 and MCP-1. Illumina Hiseq 2500 was applied to characterize the transcriptome profiles. DESeq2 was used to analyze the differential expression genes (DEGs). GOseq R package and Omicsbean were used to analyze the Gene Ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) enrichment analysis. ARPE-19 and hRPE cells were divided into control group and LPS group
and then real-time PCR was performed to verify the selected DEGs of RNA-seq results.
Results
2
The mRNA levels of IL-6
IL-8 and MCP-1 were markedly up-regulated after LPS stimulation for 24 h (
P
<
0.01). Compared with the blank control cells
a total of 2 345 DEGs were detected in LPS treated ARPE-19 cells by RNA-seq
and among them
1 594 were up-regulated and 751 were down-regulated. GO and KEGG enrichment analyses showed that LPS stimulation activated immune related pathways
Toll-like receptor
MAPK
HIF-1
TNF
PI3K-Akt
NOD-like receptor
mTOR
VEGF signal pathway
transcription factors (JUN
JUNB
SRF
ATF)
and AKT1 and SRC genes.
Conclusion
2
LPS- induced inflammation in ARPE-19 cells may be associated with the activation of PI3K-Akt-mTOR signal pathway
NOD-like receptor
VEGF signal pathway and transcription factors FOS
JUN
JUNB and SRF
as well as AKT1 and SRC genes.
关键词
Keywords
references
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