Bioinformatics study on the effect of glucocorticoid on miRNA expression in bone microvascular endothelial cells of femoral head in rats

doi:10.3877/cma.j.issn.1673-9450.2020.02.005

Abstract

Abstract:

Objective

To explore the effect of glucocorticoid on miRNA expression profile of femoral head bone microvascular endothelial cell(BMEC) and bioinformatics analysis.

Methods

Fourteen SPF 8-week-old female SD rats were selected. After 1 week of adaptive feeding, they were divided into 2 groups by random number table method, 7 in model group and 7 in control group. The model group was intraperitoneally injected with lipopolysaccharide (20 μg/kg) for two consecutive injections 24 h apart, and intramuscular injection of high-dose methylprednisolone (40 mg/kg) for three consecutive injections 24 h apart. The control group was given intraperitoneal injection and intramuscular injection of the same amount of normal saline at the same time. After 4 weeks, 2 of the rats from each group were sectioned randomly to confirm the establishment of the model. And the femoral head BMEC were cultured, identified and tested using bioinformatics methods. Such as the Agilent GeneSpring were used to identify the differentially expressed miRNA, miRWalk2.0 software were used to explore target genes, Panther database was used to predict signaling pathway of target genes, Gene ontology (GO) was used to analysis the functional annotation of target genes.

Results

The rat model of early osteonecrosis of the femoral head (ONFH) was successfully established by using lipopolysaccharide and methylamydron. The purity of BMEC was as high as 98.5%.Compared with control group, 4 miRNA (2 up-regulated miRNA: miRNA-132-3p, miRNA-335, and 2 down-regulated miRNA: miRNA-466b-2-3p, let-7c-1-3p)were detected in the model group, 3 808 target genes of which were detected. The signaling pathways regulated by target genes may include JAK/STAT signaling pathway and so on, their enrichment biological pathways include metabolic process and so on, localization in cell include organelle, nucleoid and so on, enrichment includes this in biological function include nucleic acid binding transcription factor activity and so on.

Conclusions

Glucocorticoid can induce the changes of miRNA expression profile in femoral head microvascular endothelial cells. The abnormal expression of miRNA-132-3p and miRNA-335 may play an important role in steroid-induced ONFH.

Key words: Hormones, Femur head necrosis, Endothelial cells, Biological information

Ju′an Yue, Randong Wang, Qidong Zhang, Bing Li, Qiang Sun, Wangyan Liu, Jiao Chen, Yingnan Li, Xiaozhong Guo. Bioinformatics study on the effect of glucocorticoid on miRNA expression in bone microvascular endothelial cells of femoral head in rats[J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2020, 15(02): 103-109.

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URL: https://zhssyxfzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1673-9450.2020.02.005

https://zhssyxfzz.cma-cmc.com.cn/EN/Y2020/V15/I02/103

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References 32

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数据库登陆号	miRNA	表达	FC(abs)	P值
MIMAT0000838	rno-miR-132-3p	上调	2.16	0.023
MIMAT0000575	rno-miR-335	上调	159.58	0.003
MIMAT0017286	rno-miR-466b-2-3p	下调	14.69	0.024
MIMAT0017087	rno-let-7c-1-3p	下调	8.60	0.009

数据库登陆号	miRNA	表达	FC(abs)	P值
MIMAT0000838	rno-miR-132-3p	上调	2.16	0.023
MIMAT0000575	rno-miR-335	上调	159.58	0.003
MIMAT0017286	rno-miR-466b-2-3p	下调	14.69	0.024
MIMAT0017087	rno-let-7c-1-3p	下调	8.60	0.009

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