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通过微阵列和生物信息学分析研究绝经后骨质疏松症患者骨髓间充质干细胞外泌体中 lncRNA 的表达谱分析。

Expression profile analysis of lncRNA in bone marrow mesenchymal stem cells exosomes of postmenopausal osteoporosis patients through microarray and bioinformatics analyses.

机构信息

Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua, China.

Affiliated Jinhua Hospital, Zhejiang University School of Medicine, Jinhua, China.

出版信息

Pathol Res Pract. 2022 Aug;236:153985. doi: 10.1016/j.prp.2022.153985. Epub 2022 Jun 17.

Abstract

BACKGROUND

Postmenopausal osteoporosis (PMOP) is the most common bone metabolic disease affecting women worldwide. In this study, we investigate the role of long non-coding RNA (lncRNA) expression in exosomes obtained from bone marrow mesenchymal stem cells (BMSCs) of patients with PMOP.

METHODS

BMSCs from patients diagnosed with PMOP and healthy post-menopausal females as controls were isolated and cultured before exosome extraction. RNA microarray technology was used to identify differentially expressed lncRNAs in exosomes from BMSCs. Bioinformatics technology was utilized to analyze the roles of differentially expressed lncRNAs. Further, RT-qPCR was used to validate differentially expressed lncRNAs in 20 pairs of clinical samples.

RESULTS

A total of 286 differentially expressed lncRNAs were detected in the exosomes from BMSCs unlike in the control group, among which 148 were up-regulated, whereas 138 were down-regulated. RT-qPCR identified five critical lncRNAs, including ENST00000593078, NR_120593, ENST00000422343, MEG3 and NR_029192. This was consistent with the microarray results and with a significant difference (P < 0.01). Based on the differentially expressed lncRNAs, we constructed lncRNA-miRNA-mRNA interaction networks. Functional analysis revealed that differentially expressed lncRNAs in patients with PMOP potentially target Wnt/β-catenin, MAPK, and PI3K-Akt pathways.

CONCLUSION

In summary, we detected several dysregulated lncRNAs regulating PMOP progression in exosomes extracted from BMSCs of affected patients acting as novel biomarkers. This in turn provides valuable data for targeted treatment of PMOP.

SUBJECTS

Genomics; Molecular biology; Orthopedics; Women's Health.

摘要

背景

绝经后骨质疏松症(PMOP)是全球范围内影响女性最常见的骨骼代谢疾病。在这项研究中,我们研究了长链非编码 RNA(lncRNA)在 PMOP 患者骨髓间充质干细胞(BMSC)来源的外泌体中的表达作用。

方法

分离和培养 PMOP 患者和健康绝经后女性的 BMSC 以提取外泌体。使用 RNA 微阵列技术鉴定 BMSC 来源的外泌体中差异表达的 lncRNA。利用生物信息学技术分析差异表达 lncRNA 的作用。进一步使用 RT-qPCR 验证 20 对临床样本中差异表达的 lncRNA。

结果

与对照组相比,BMSC 来源的外泌体中检测到 286 个差异表达的 lncRNA,其中 148 个上调,138 个下调。RT-qPCR 鉴定出五个关键 lncRNA,包括 ENST00000593078、NR_120593、ENST00000422343、MEG3 和 NR_029192。这与微阵列结果一致,且差异有统计学意义(P < 0.01)。基于差异表达的 lncRNA,我们构建了 lncRNA-miRNA-mRNA 相互作用网络。功能分析显示,PMOP 患者中差异表达的 lncRNA 可能靶向 Wnt/β-catenin、MAPK 和 PI3K-Akt 通路。

结论

总之,我们在受影响患者的 BMSC 提取的外泌体中检测到了几个调节 PMOP 进展的失调 lncRNA,它们作为新的生物标志物具有重要意义。这反过来为 PMOP 的靶向治疗提供了有价值的数据。

研究对象

基因组学;分子生物学;骨科;妇女健康。

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