Project description:Objective: To explore the molecular mechanism of Codonopsis pilosula in enhancing the immune function of aging mice, the expression profile of long-chain non-coding RNA (lncRNA), mRNA in spleen of mice was detected. Methods: An aging mouse model was established, the aging mice were treated with low, middle and high doses of Codonopsis pilosula, and the aging related indexes, spleen pathology and cellular ultrastructure were detected and analyzed. Then microarray analysis and bioinformatics methods were used to analyze the lncRNA and mRNA expression profile. GO enrichment analysis and KEGG pathway analysis were performed to analyze the differential genes between groups using KOBAS. Finally, the common lncRNAs and mRNAs in high-dose Codonopsis pilosula treatment group compared with the model group and the model group compared with the control group were selected for lncRNA-mRNA co-expression network construction, and real-time quantitative polymerase chain reaction (qRT-PCR) was used to verify the genes in the network. Results: The aging mouse model was successfully established. Codonopsis pilosula can improve the pathology and cellular ultrastructure of spleen in aging mice. Microarray analysis showed that 96 lncRNAs and 65 mRNAs changed significantly after aging, 623 lncRNAs and 435 mRNAs expressed abnormally after high-dose Codonopsis pilosula treatment. KEGG pathways involved in aging and high-dose Codonopsis pilosula treatment are mainly related to immunity, including allograft rejection, graft-versus-host disease, autoimmune thyroid disease, antigen processing and presentation and so on. The three mRNAs (Enpp6, Cped1, and Galnt15) in lncRNA-mRNA co-expression network may be related to Codonopsis pilosula improving the immunity of aging mice, which were verified by qRT-PCR. Conclusion: Codonopsis pilosula has a certain protective effect on the spleen of aging mice and the lncRNA-mRNA network may play an important role in enhancing the immunity of aging mice.

Project description:BACKGROUND: Competitive endogenous RNA (ceRNA) reveals new mechanisms for interactions between RNAs. However, knowledge of ceRNA regulatory networks in macrophages infected with Talaromyces marneffei (T. marneffei, TM) is still limited. This study aims to explore the expression profiles of lncRNA, miRNA and mRNA, and the changes of ceRNA network related to immunity, inflammation, metabolism in TM-infected macrophage. METHODS: Next-generation sequencing technology (NGS) was used to obtain mRNA, miRNA and lncRNA expression profiles in TM-infected macrophages and normal macrophages. Cuffdiff and DESeq2 software packages are used to identify differentially expressed lncRNA, miRNA and mRNA. The function enrichment analysis was performed by GOseq package in R platform, and the lncRNA–miRNA–mRNA interaction ceRNA network was established in Cytoscape. Functional enrichment analysis was performed on genes contained in the ceRNA network, and genes significantly enriched in functional pathways were selected for qRT-PCR verification. RESULTS: A total of 119 lncRNAs, 28 miRNAs and 208 mRNAs were identified as differentially expressed RNAs in TM-infected macrophages. Among them, 38 lncRNAs, 10 miRNAs and 109 mRNAs are contained in the ceRNA regulatory network. GO analysis and KEGG pathway of mRNA in the ceRNA network showed that macrophages infected with TM activated pathways and functions related to immune response, metabolic response and inflammatory response. Select genes that are significantly abundant in GO analysis and KEGG pathway. The core genes were selected for quantitative real-time PCR (qRT-PCR) to verify consistency with the sequencing results. Eight groups of ceRNAs consisting of 3 mRNAs, 5 lncRNAs, and 2 miRNAs were verified. They are mRNA CSF3, IL24, LIF. lncRNA IL6R-AS1, LINC02009, AC068831.1, AC006252.1, LINC02466. These genes may be involved in immune response and inflammatory response-related pathways and functions in TM-infected macrophages. CONCLUSIONS: The CeRNA network plays an important role in understanding the mechanism of TM infection in macrophages. This study may provide effective and novel insights for further understanding the underlying mechanism of Talaromyces marneffei.

Dataset Information

Integrative analysis of lncRNAs, miRNAs, and mRNA-associated ceRNA network in lung tissue of aging mice and changes after intervention of Codonopsis pilosula (miRNA dataset)

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