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Differentially expressed lncRNAs and mRNAs in primary human airway granulation fibroblasts before and after β-elemene treatment for 48h

ABSTRACT: There is an urgent need to identify effective medications to inhibit the abnormal airway granulation fibroblast activity, which plays a critical role in granulation tissue hyperplasia and scar formation during wound-healing after endobronchial intervention in the human airway, so as to prevent benign airway restenosis. Our previous study shown that β-elemene, a widely used effective ingredient of traditional Chinese medicine, effectively inhibited proliferation and survival of primary human airway granulation fibroblasts (PHAGF) in vitro, and promotes granulation tissue degeneration in vivo. To further investigate the mechanistic basis of these effects, we have employed LncRNA Microarray expression profiling as a discovery platform to identify differentially expressed lncRNAs and mRNAs in β-Elemene-treated PHAGF, thus enriching β-Elemene-altered pathway associated with cell proliferation and apoptosis, and thereby constructing a β-elemene-altered competitive endogenous RNAs (ceRNA) network that modulates this pathway. The results revealed ILK/Akt pathway, containing the upstream factor, ILK, the downstream cell cycle gene, Cyclin D1, and the anti-apoptosis gene, Bcl-2, were suppressed in β-Elemene-treated PHAGF. LncRNA-MIR143HG, which acts as a miR-1275 ceRNA to modulate ILK expression was also suppressed. Furhter validation via wet-lab experiments revealed that β-Elemene down-regulated LncRNA MIR143HG in PHAGF to reduce the competing sponging effect of MIR143HG on microRNA miR-1275, which targets upstream gene ILK in ILK/Akt pathway, thereby inducing the fibroblasts cell cycle arrest and apoptosis by suppressing ILK/Akt pathway.

ORGANISM(S): Homo sapiens

PROVIDER: GSE155297 | GEO | 2021/06/16

REPOSITORIES: GEO

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Project description:Background Rotator cuff tears (RCT) is a common musculoskeletal disorder in the shoulder which cause pain and functional disability. Diabetes mellitus (DM) is characterized by impaired ability of producing or responding to insulin and has been reported to act as a risk factor of the progression of rotator cuff tendinopathy and tear. Long non-coding RNAs (lncRNAs) are involved in the development of various diseases, but little is known about their potential roles involved in RCT of diabetic patients. Methods RNA-Sequencing (RNA-Seq) was used in this study to profile differentially expressed lncRNAs and mRNAs in RCT samples between 3 diabetic and 3 nondiabetic patients. Gene ontology (GO) and Kyoto encyclopedia of genes and genomes (KEGG) pathway analysis were performed to annotate the function of the differentially expressed genes (DEGs). LncRNA-mRNA co-expression network and competing endogenous RNA (ceRNA) network were constructed to elucidate the potential molecular mechanisms of DM affecting RCT. Results In total, 505 lncRNAs and 388 mRNAs were detected to be differentially expressed in RCT samples between diabetic and nondiabetic patients. GO functional analysis indicated that related lncRNAs and mRNAs were involved in metabolic process, immune system process and others. KEGG pathway analysis indicated that related mRNAs were involved in ferroptosis, PI3K-Akt signaling pathway, Wnt signaling pathway, JAK-STAT signaling pathway and IL-17 signaling pathway and others. LncRNA-mRNA co-expression network was constructed, and ceRNA network showed the interaction of differentially expressed RNAs, comprising 5 lncRNAs, 2 mRNAs, and 142 miRNAs. TF regulation analysis revealed that STAT affected the progression of RCT by regulating the apoptosis pathway in diabetic patients. Conclusions We preliminarily dissected the differential expression profile of lncRNAs and mRNAs in torn rotator cuff tendon between diabetic and nondiabetic patients. And the bioinformatic analysis suggested some important RNAs and signaling pathways regarding inflammation and apoptosis were involved in diabetic RCT. Our findings offer a new perspective on the association between DM and progression of RCT.

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Differentially expressed lncRNAs and mRNAs in primary human airway granulation fibroblasts before and after β-elemene treatment for 48h

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