RESUMO
Objective: We have already demonstrated that mesenchymal stem cells from patients with ankylosing spondylitis (ASMSCs) exhibited greater adipogenic differentiation potential than those from healthy donors (HDMSCs). Here, we further investigated the expression profile of long noncoding RNA (lncRNA) and mRNA, aiming to explore the underlying mechanism of abnormal adipogenic differentiation in ASMSCs. Methods: HDMSCs and ASMSCs were separately isolated and induced with adipogenic differentiation medium for 10 days. Thereafter, lncRNAs and mRNAs that were differentially expressed (DE) between HDMSCs and ASMSCs were identified via high-throughput sequencing and confirmed by quantitative real-time PCR (qRT-PCR) assays. Then, the DE genes were annotated and enriched by GO analysis. In addition, protein interaction network was constructed to evaluate the interactions between DE mRNAs and to find hub nodes and study cliques. Besides, co-expression network analysis was carried out to assess the co-expressions between DE mRNA and DE lncRNAs, and competing endogenous RNA (ceRNA) network analysis were conducted to predict the relationships among lncRNAs, mRNAs and miRNAs. The signaling pathways based on the DE genes and the predicted DE genes were enriched by KEGG analysis. Results: A total of 263 DE lncRNAs and 1376 DE mRNAs were found during adipogenesis in ASMSCs. qRT-PCR indicated that the expression of the top 20 mRNAs and the top 10 lncRNAs was consistent with the high-throughput sequencing data. Several lncRNAs (NR_125386.1, NR_046473.1 and NR_038937.1) and their target genes (SPN and OR1AIP2), together with the significantly co-expressed pairs of DE lncRNAs and DE mRNAs (SLC38A5-ENST00000429588.1, TMEM61-ENST00000400755.3 and C5orf46-ENST00000512300.1), were closely related to the enhanced adipogenesis of ASMSCs by modulating the PPAR signaling pathway. Conclusion: Our study analyzed the expression profiles of DE lncRNAs and DE mRNAs during adipogenesis in ASMSCs and HDMSCs. Several DE lncRNAs, DE mRNAs and signaling pathways that probably participate in the aberrant adipogenesis of ASMSCs were selected for future study. These results will likely provide potential targets for our intervention on fat metaplasia and subsequent new bone formation in patients with AS in the future.
RESUMO
BACKGROUND: Little is known about the effect of different rearing conditions on the effects of methamphetamine and whether the introduction of enriched rearing conditions at different stages of development could produce different behavioral outcomes. METHODS: In Experiment 1, rats were reared in either enriched (EE) or isolated environments (IE) from PND 21 to 60. In Experiment 2, two groups of animals were handled in the same fashion as those in Experiment 1. Additional two groups were housed in IE during the first 20 or 30 days and then housed under EE for the remaining 20 or 10 days respectively. Locomotor activity and Morris Water Maze were tested. The effects of rearing conditions on methamphetamine (METH) self-administration were investigated. RESULTS: IE animals exhibited higher levels of locomotion than EE animals, but EE animals showed enhanced Morris water maze performance. Animals reared in IE for 30 and 40 days more readily acquired METH self-administration, compared to those reared in IE for 20 and in EE for 40 days respectively. However, the effect of rearing conditions was only seen at the lowest dose tested under FR schedule and breakpoints obtained from PR schedule were not significantly affected. Those reared in IE for 20 and EE for 40 days animals produced significantly fewer responses during the extinction and cue-induced reinstatement of METH self-administration, compared with animals reared in IE for 30 and 40 days, respectively. CONCLUSION: Rearing condition plays a significant role in locomotor activity, spatial memory and behavioral effects of METH.