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唐暹, 周丹丹, 尹林玉, 佘艳, 刘松涛, 艾坤.基于PBMCs转录组学探讨抗阻运动改善绝经后骨质疏松症患者骨代谢的作用机制[J].湖南中医药大学学报,2026,46(5):964-974[点击复制] |
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| 基于PBMCs转录组学探讨抗阻运动改善绝经后骨质疏松症患者骨代谢的作用机制 |
| 唐暹,周丹丹,尹林玉,佘艳,刘松涛,艾坤 |
| (湖南中医药大学, 湖南 长沙 410208;湘南学院附属医院, 湖南 郴州 423000) |
| 摘要: |
| 目的 基于外周血单核细胞（PBMCs）转录组学分析，探讨3个月中等强度抗阻运动改善绝经后骨质疏松症（PMOP）患者骨代谢的潜在作用机制。方法 选取2025年4月至2025年8月在湘南学院附属医院康复医学科门诊就诊的16例PMOP患者，随机分为对照组与运动组（各8例）。对照组给予碳酸钙D3补充及健康宣教；运动组在对照组基础上进行中等强度抗阻运动干预，每周3次，每次60 min，持续3个月。采用ELISA检测干预前后血清Ⅰ型前胶原羧基端原肽（PICP）、Ⅰ型胶原交联C端肽（CTX）含量。分离干预结束后采集的血液中的PBMCs进行转录组测序；采用edgeR对mRNA、lncRNA和miRNA进行差异表达分析；使用STRING数据库构建PPI网络并筛选枢纽基因；对基因集进行GO和KEGG富集分析；整合差异表达分析、数据库靶向预测与共表达分析，构建ceRNA网络；采用单样本基因集富集分析（ssGSEA）进行免疫浸润分析；RT-qPCR验证关键基因表达水平。结果 干预前，两组患者血清PICP和CTX含量比较，差异均无统计学意义（P>0.05）。与干预前比较，干预后运动组血清PICP水平升高（P＜0.01）。与对照组比较，运动组干预后血清PICP含量升高（P<0.05）。与对照组比较，运动组干预前后血清PICP含量的差值升高（P<0.05），干预前后血清CTX的差值降低（P<0.05）。差异表达分析共识别出529个mRNA、500个lncRNA、68个miRNA；PPI网络识别出10个枢纽基因；GO和KEGG富集分析显示，差异基因主要涉及吞噬作用/吞噬体、髓系白细胞激活、白细胞介素-6调控、C型凝集素受体信号通路、Toll样受体（TLR）信号通路等先天免疫与炎症信号，并与过氧化物酶体增殖物激活受体、促分裂原激活蛋白激酶、腺苷酸活化蛋白激酶及甲状旁腺激素等相关通路相关。ceRNA网络包括2个lncRNA、5个miRNA和7个mRNA，其中lncRNA核富集丰度转录本1（NEAT1）与母源表达基因3（MEG3）为ceRNA网络关键节点，mRNA富集于机械刺激/肌肉牵张反应、破骨细胞分化的调控、脂多糖介导信号、TLR与钙信号及钙再吸收相关通路。免疫浸润分析显示，运动组中性粒细胞、巨噬细胞、树突状细胞及髓源性抑制性细胞等免疫细胞的浸润评分降低（P＜0.05），且ceRNA网络mRNA与多类先天免疫细胞浸润呈正相关（P＜0.05）。TLR4与AP-1转录因子亚单位（FOS）作为PPI网络与ceRNA网络的共同关键基因，经RT-qPCR验证在运动组表达下调（P＜0.05）。结论 3个月中等强度抗阻运动可促进PMOP患者骨代谢良性趋势改变，其机制可能与先天免疫功能状态重塑、炎症信号抑制及钙/内分泌相关通路调控有关。NEAT1与MEG3介导的ceRNA网络可能通过调控miRNA影响TLR4、FOS等关键基因表达，从而参与运动介导的免疫炎症改变与骨代谢调控。 |
| 关键词: 绝经后骨质疏松症|抗阻运动|外周血单核细胞|转录组学|ceRNA|lncRNA|miRNA|免疫浸润 |
| DOI：10.3969/j.issn.1674-070X.2026.05.013 |
| 投稿时间：2026-02-11 |
| 基金项目:湖南省自然科学基金项目（2024JJ7533）；湖南中医药大学校级科研项目（2024XJZC003）；湖南中医药大学研究生科研创新项目（2024CX001）。 |
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| Mechanism of action of resistance exercise in improving bone metabolism in postmenopausal osteoporosis based on PBMCs transcriptomics |
| TANG Xian, ZHOU Dandan, YIN Linyu, SHE Yan, LIU Songtao, AI Kun |
| (Hunan University of Chinese Medicine, Changsha, Hunan 410208, China;Affiliated Hospital of Xiangnan University, Chenzhou, Hunan 423000, China) |
| Abstract: |
| Objective To explore the potential mechanism of action of 3-month moderate-intensity resistance exercise in improving bone metabolism in patients with postmenopausal osteoporosis (PMOP) based on peripheral blood mononuclear cells (PBMCs) transcriptomic analysis. Methods Sixteen PMOP patients who visited the outpatient clinic of the Department of Rehabilitation Medicine, Affiliated Hospital of Xiangnan University from April 2025 to August 2025 were enrolled and randomly divided into control group and exercise group, with 8 patients in each group. The control group received calcium carbonate D3 supplementation and health education, while the exercise group received moderate-intensity resistance exercise intervention on the basis of the control group, 3 times per week, 60 min per session, for 3 months. Serum levels of type I procollagen carboxy-terminal propeptide (PICP) and C-terminal telopeptide of type I collagen (CTX) were measured using ELISA before and after the intervention. PBMCs were isolated from post-intervention blood for transcriptome sequencing. Differential expression analysis of mRNAs, lncRNAs, and miRNAs was performed using edgeR. The STRING database was used to construct a protein-protein interaction (PPI) network and screen hub genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed on gene sets. A competing endogenous RNA (ceRNA) network was constructed by integrating differential expression analysis, database-based target prediction, and co-expression analysis. Immune infiltration analysis was performed using single-sample gene set enrichment analysis (ssGSEA). Key gene expression levels were validated using reverse transcription quantitative real-time PCR (RT-qPCR). Results Before the intervention, there were no statistically significant differences in serum PICP and CTX levels between the two groups (P>0.05). Compared with before intervention, serum PICP levels in the exercise group increased after intervention (P<0.01). Compared with the control group, serum PICP levels in the exercise group were higher after intervention (P<0.05). Compared with the control group, the exercise group showed a greater increase in the difference in serum PICP levels before and after intervention (P<0.05), and a greater decrease in the difference in serum CTX levels (P<0.05). Differential expression analysis identified 529 mRNAs, 500 lncRNAs, and 68 miRNAs. The PPI network identified 10 hub genes. GO and KEGG enrichment analyses showed that the differentially expressed genes were mainly involved in innate immune and inflammatory signaling, including phagocytosis/phagosome, myeloid leukocyte activation, interleukin-6 (IL-6) regulation, C-type lectin receptor (CLR) signaling pathway, and Toll-like receptor (TLR) signaling pathway, and were also associated with pathways related to peroxisome proliferator-activated receptor (PPAR), mitogen-activated protein kinase (MAPK), AMP-activated protein kinase (AMPK), and parathyroid hormone (PTH). The ceRNA network included 2 lncRNAs, 5 miRNAs, and 7 mRNAs. Among them, lncRNA nuclear-enriched abundant transcript 1 (NEAT1) and maternally expressed gene 3 (MEG3) were key nodes in the ceRNA network. The mRNAs in this network were enriched in pathways related to mechanical stimulation/muscle stretch response, regulation of osteoclast differentiation, lipopolysaccharide-mediated signaling, TLR and calcium signaling, and calcium reabsorption. Immune infiltration analysis showed that infiltration scores of neutrophils, macrophages, dendritic cells, and myeloid-derived suppressor cells were decreased in the exercise group (P<0.05), and the mRNAs in the ceRNA network were positively correlated with the infiltration of multiple innate immune cells (P<0.05). TLR4 and AP-1 transcription factor subunit (FOS), as common key genes in both the PPI and ceRNA networks, were downregulated in the exercise group as verified by RT-qPCR (P<0.05). Conclusion Three months of moderate-intensity resistance exercise can promote a favorable shift in bone metabolism in PMOP patients, and the underlying mechanisms may be associated with the remodeling of innate immune functional status, inhibition of inflammatory signaling, and regulation of calcium/endocrine-related pathways. The NEAT1- and MEG3-mediated ceRNA network may modulate the expression of key genes such as TLR4 and FOS through miRNA regulation, thereby participating in exercise-mediated immunoinflammatory changes and bone metabolism regulation. |
| Key words: postmenopausal osteoporosis|resistance exercise|peripheral blood mononuclear cell|transcriptomics|competing endogenous RNA|lncRNA|miRNA|immune infiltration |
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