Network construction of key modulatory miRNA and downstream target genes in the progression of osteoporosis under simulated microgravity through high-throughput sequencing
Journal Title: Journal of Air Force Medical University - Year 2023, Vol 44, Issue 6
Abstract
Objective To screen the differential expression profiles of miRNAs in normal and weightless osteoporosis cell samples by using high-throughput sequencing technology, and to analyze the biological information of the differentially expressed miRNAs (DEmiRNAs), so as to explore the pathogenic mechanism of weightless osteoporosis from the molecular level. Methods Mouse bone marrow mesenchymal stem cell lines were divided into Ground group and simulated microgravity ( SMG) group, both groups were treated with BMP-2 for osteogenic differentiation induction, and the SMG group was put into a clinostat at 30 r / min for 48 h to simulate weightless conditions. Then, total RNA was extracted from the samples of the two groups, and transcriptome sequencing was performed by Illumina NovaSeq 6000. DEseq2 algorithm was used and the threshold was set as fold change > 2 and P < 0. 05 to screen and analyze the differentially expressed genes (DEGs) of the cell samples of the Ground group and SMG group. Hierarchical clustering and volcano map analysis were used to demonstrate DEmiRNAs, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to understand the molecular functions and signaling pathways enriched by differential genes, and the involvement of mRNA in the biological processes of the proliferation and differentiation of osteoblasts and osteoclasts was further explored. Results By comparing the Ground group and SMG group, a total of 44 DEmiRNAs were identified, including 16 up-regulated miRNAs and 28 down-regulated miRNAs. GO analysis showed that DEGs were enriched in the biological processes of osteoblasts and osteoclasts, and were involved in cell proliferation, differentiation, apoptosis and other biological processes. KEGG results showed that DEGs were significantly enriched in TNF and FoxO signaling pathways. Conclusion DEmiRNAs in weightless osteoporosis screened by Illumina high-throughput sequencing have significant differences in their functions, and it was found that these DEmiRNAs may be involved in various molecular biological processes such as osteoblasts and osteoclasts in weightless osteoporosis. The results of this study provide a theoretical basis and new ideas for further research on the pathogenesis and clinical treatment of weightless osteoporosis.
Authors and Affiliations
LI Weihang, ZHAO Yingjing, SHI Quan, WANG Dong, DING Ziyi, GAO Bo, YAN Ming, LUO Zhuojing
Keywords
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