韩书娜1，孙一2，叶琼3
(1.四川外国语大学 体育部，重庆 400033；2.吉林大学 体育学院，吉林 长春 130012；3.重庆幼儿师范高等专科学校 体育教研室，重庆 404000)
摘 要：观察规律抗阻训练对老年男性循环内皮祖细胞功能的影响并探讨miR-21-5p/血小板反应蛋白(TSP-1)在其间的可能作用机理。60名健康、无规律运动习惯的老年男性随机分为对照组和运动组，对照组维持日常生活习惯，运动组进行3次/周、共12周的抗阻训练干预。分别于试验前后，利用超声诊断系统测定肱动脉血流介导的血管舒张功能(FMD)；取外周血分离培养内皮祖细胞，利用β-半乳糖苷酶染色法检测衰老水平，MTT比色法、Matrigel管腔形成实验分别检测体外增殖和成管能力，通过裸鼠颈动脉内膜拉脱模型检测在体内皮损伤修复能力，实时荧光定量PCR检测miR-21-5p和TSP-1 mRNA表达量，免疫印迹法测定TSP-1蛋白表达量；通过脂质体转染分别过表达miR-21-5p(miR-21-5p模拟物)和抑制TSP-1(TSP-1 siRNA)表达后，检测内皮祖细胞衰老和功能。结果：(1)训练依从性和安全性：运动组1例(3.3%)脱落，训练计划完成率为96.8%，无严重不良反应事件或心血管相关事件发生。(2)血管内皮功能和内皮祖细胞功能：试验后，与对照组比较，运动组FMD升高(P<0.05)，内皮祖细胞β-半乳糖苷酶阳性细胞下降(P<0.05)，体外增殖和成管能力增加(P<0.05)，体内损伤血管再内皮化功能提高(P<0.05)，miR-21-5p表达上调(P<0.05)，TSP-1 mRNA和蛋白表达下调(P<0.05)。(3)细胞转染试验：转染miR-21-5p模拟物诱导miR-21-5p过表达后，内皮祖细胞TSP-1 mRNA和蛋白表达下降(P<0.05)，体外增殖、成管能力以及体内损伤血管再内皮化功能改善(P<0.05)，然而，β-半乳糖苷酶阳性细胞无显著性变化(P>0.05)；转染TSP-1 siRNA诱导TSP-1沉默后，β-半乳糖苷酶阳性细胞下降(P<0.05)，体外增殖、成管能力以及体内损伤血管再内皮化功能提升(P<0.05)。结论：抗阻训练通过上调miR-21-5p抑制TSP-1表达来改善老年男性内皮祖细胞功能，通过下调TSP-1延缓内皮祖细胞衰老，因此miR-21-5p和TSP-1是抗阻训练发挥心血管保护效应的重要靶点。
关 键 词：运动生物化学；抗阻训练；内皮祖细胞；血管内皮功能；老年男性
中图分类号：G804.7 文献标志码：A 文章编号：1006-7116(2025)01-0150-07
A study on resistance training improves age-related circulating endothelial progenitor cell functional impairment in old males by miR-21- 5p/TSP-1 pathway
HAN Shuna1，SUN Yi2，YE Qiong3
(1.Department of Physical Education，Sichuan International Studies University，Chongqing 400033，China；2.School of Physical Education，Jilin University，Changchun 130012，China；3.Department of Physical Education，Chongqing Preschool Education College，Chongqing 404000，China)
Abstract: To observe the effect of regular resistance training on the function of circulating endothelial progenitor cells in elderly men and to explore the possible mechanism of miR-21-5p/thrombospondin 1 (TSP-1). 60 healthy old males with no regular exercise habits were randomly divided into control group and exercise group. The control group maintained their daily habits, and the exercise group underwent resistance training intervention three times per week for a total of 12 weeks. Before and after the experiment, an ultrasonic diagnostic system was used to measure brachial artery flow mediated dilation (FMD). Peripheral blood was taken to isolate and culture endothelial progenitor cells, β-galactosidase staining to detect the aging level, MTT colorimetry and Matrigel tube formation assay to detect the proliferation and tube-forming ability in vitro respectively, the repair ability of endothelial damage in vivo through the nude mouse carotid artery intimal pull-off model, real-time fluorescence quantitative PCR to detect the expression of miR-21-5p and TSP-1 mRNA, and the expression of TSP-1 protein by Western blotting, were determined. After overexpressing miR-21-5p (miR-21-5p mimic) and inhibiting TSP-1 (TSP-1 siRNA) expression respectively by lipofectamine transfection, endothelial progenitor cell senescence and function were detected. The results show that: (1) Training compliance and safety: 1 case (3.3%) dropped out, the training plan completion rate was 96.8%, and no serious adverse events or cardiovascular-related events occurred in the exercise group. (2) Vascular endothelial function and endothelial progenitor cell function: After the experiment, compared with the control group, FMD increased (P<0.05), β-galactosidase-positive cells of endothelial progenitor cells decreased (P<0.05), the proliferation and tube formation abilities in vitro were increased (P<0.05), the reendothelialization function of injured blood in vivo was improved (P<0.05), the expression of miR-21-5p was up-regulated (P<0.05), and the expression of TSP-1 mRNA and protein was down-regulated (P<0.05) in the exercise group. (3) Cell transfection test: After transfection with miR-21-5p mimic to induce the overexpression of miR-21-5p, the expression of TSP-1 mRNA and protein in endothelial progenitor cells decreased (P<0.05), the proliferation and tube formation ability in vitro and reendothelialization function of injured blood in vivo were improved (P<0.05), however, there were no significant changes in β-galactosidase-positive cells (P>0.05). After transfection of TSP-1 siRNA to induce TSP-1 silencing, β-galactosidase-positive cells decreased (P<0.05), the proliferation and tube formation ability in vitro and reendothelialization function of injured blood in vivo increased (P<0.05). The conclusion reveal that: resistance training inhibits TSP-1 expression by up-regulating miR-21-5p, improves endothelial progenitor cell function, and delays endothelial progenitor cell aging by down-regulating TSP-1 in elderly men. Therefore, miR-21-5p and TSP-1 are important targets for the cardiovascular protective effects of resistance training
Keywords: sports biochemistry；resistance training；endothelial progenitor cells；vascular endothelial function；old males