牛膝醇提取物含药血清促进骨髓间充质干细胞增殖的关键microRNAs研究*

作者：钟秋辉1,2，周宇昊1,2，马笃军1,2，彭力平1，毕玉杰1,2，王立新1,2，文胤翔2

单位：1.广州中医药大学第四临床医学院，广东 深圳 518033； 2.广州中医药大学，广东 广州 510006

引用：引用：钟秋辉，周宇昊，马笃军，彭力平，毕玉杰，王立新，文胤翔.牛膝醇提取物含药血清促进骨髓间充质干细胞增殖的关键microRNAs研究[J].中医药导报,2025,31(8):58-65,85.

DOI：10.13862/j.cn43-1446/r.2025.08.010

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摘要：

目的：探讨含有牛膝（Achyranthes bidentata）醇提取物（AEAB）的兔血清影响骨髓间充质干细胞（BMMSC）增殖的关键微小RNA（miRNAs）。方法：BMMSC暴露于3种不同剂量的AEAB含药血清中（低剂量，每天1 g/kg；中剂量，每天3 g/kg；高剂量，每天6 g/kg），并检测其活力。对AEAB含药血清中剂量组和模拟组的BMMSC进行miRNA测序，以鉴定组间差异表达的miRNA（DEMs）。然后，探索miRNA-mRNA的关系，并进行功能富集分析和转录因子（TFs）-miRNA-mRNA相互作用网络分析。最后，通过实验验证关键DEMs在BMMSC中的表达和功能。结果：3种不同剂量的AEAB兔血清均可以显著提高BMMSC的活力。通过miRNA测序，在AEAB含药血清中剂量组和模拟组中共鉴定出40个DEMs。然后，149种miRNA-mRNA相互作用被鉴定出来，例如miR-669o-5p-Gnal。功能富集分析发现，包括miR-1946a在内的DEMs主要参与细胞-细胞黏附介质的活性。此外，33种miRNA-TF相互作用被鉴定出来，例如miR-23b-5p-Myc。最后，qPCR分析证实miR-1946a在AEAB含药血清处理后表达下调，miR-669o-5p和miR-23b-5p表达上调，与miRNA测序分析结果一致。抑制miR-669o-5p和miR-23b-5p后，含AEAB的血清对BMMSC活性的影响被抑制。结论：AEAB含药血清可能通过上调miR-23b-5p和miR-669o-5p促进BMMSC的增殖。此外，miR-1946a下调可能通过调控细胞-细胞黏附介质活性介导AEAB含药血清对BMMSC的促增殖作用。

关键词：骨质疏松症；牛膝醇提取物；骨髓间充质干细胞；差异表达miRNAs；功能富集分析

Abstract：

Objective: To explore key microRNAs (miRNAs) by which rabbit serum containing alcoholic extract of Achyranthes bidentata (AEAB) affected proliferation of bone marrow mesenchymal stem cells (BMSCs). Methods: BMSCs were exposed to three doses of AEAB-containing serum (low dose,1 g/kg; medium dose, 3 g/kg; and high dose, 6 g/kg), and their viability was detected. miRNA sequencing for the medium dose and mock (negative control) groups was performed to identify differentially expressed miRNAs (DEMs) between groups. miRNA-mRNA relationships were then explored, followed by functional enrichment analysis and analysis of the transcription factors (TFs)-miRNA-mRNA interaction network. Finally, the expression and function of key DEMs were experimentally verified in BMSCs. Results: BMSC viability was significantly increased after treatment with three doses of AEAB-containing serum. Totally 40 DEMs were identified in the medium dose group and mock group through miRNA sequencing. Then, 149 miRNA-mRNA interactions were identified, such as miR-669o-5p-G Protein Subunit Alpha L (Gnal). Functional enrichment analysis showed that the DEMs including miR-1946a were primarily involved in cell-cell adhesion mediator activity. Furthermore, 33 miRNA-TF interactions, such as miR-23b-5p-Myc. Finally, qPCR analysis confirmed down-regulated expression of miR-1946a and up-regulated expression of miR-669o-5p and miR-23b-5p after treatment with AEAB-containing serum, which was consistent with the miRNA sequencing analysis results. The effect of AEAB-containing serum on BMSC activity was inhibited after inhibition of miR-669o-5p and miR-23b-5p. Conclusions: The AEAB-containing serum may promote BMSC proliferation via up-regulating miR-23b-5p and miR-669o-5p. Moreover, down-regulation of miR-1946a may mediate the effect of AEAB-containing serum on BMSC proliferation by mediating cell-cell adhesion mediator activity.

Key words：osteoporosis; alcoholic extract of Achyranthes bidentata; bone marrow mesenchymal stem cells; differentially expressed miRNAs; functional enrichment analysis

发布时间：2026-01-06

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