运动训练对尿源干细胞原代产量及早期生物学特性的影响

doi:10.3969/j.issn.1004-616x.2025.06.008

摘要/Abstract

摘要： 目的：评估运动训练对尿源干细胞(USCs)原代产量及早期生物学特性的影响，并考察晚间固定时段采样的可行性，为无创、标准化获取USCs提供实验依据。方法：通过设计自身对照，即同一志愿者于晨醒后(6:00—7:00)、上午(9:00—10:00)、下午 (14:00—15:00)、晚间(20:00—21:00)4个时点留尿，通过比较原代克隆形成数以确定最佳采样时段；在最佳采样时段内，先静息 7 d，再连续7 d进行固定强度跑步(2.8 km，16~18 min)，运动后即刻留尿。两组尿液按统一流程分离培养USCs。通过形态学观察评估运动训练对USCs细胞形态的影响，采用四甲基噻唑蓝(MTT)法检测USCs生长活力并绘制7 d生长曲线，采用逆转录实时荧光定量PCR(RT-qPCR)法检测USCs干性基因OCT4、C-MYC、SOX2、NANOG mRNA的表达水平，Western blot法检测OCT4、C-MYC蛋白的表达。结果：与其余时段相比，晚间(20:00-21:00)采样可稳定获得USCs克隆，克隆形成率更高。在晚间时段内，与静息对照组相比，运动组的USCs细胞形态更为均一，胞质更透亮，细胞间连接更紧密；细胞活力增殖曲线第7天D(570)显著升高(P<0.01)，表现出更持久的增殖活力；干性基因OCT4、C-MYC mRNA的表达水平上调(P<0.05或P<0.01)，SOX2、NANOG mRNA表达水平无显著差异(P>0.05)；OCT4、C-MYC的蛋白表达水平上调(P<0.01)。结论：晚间(20:00-21:00)取材联合运动训练可显著提升USCs提取效率并优化其早期生物学特性，为后续的规模化扩增与临床转化提供了简单、可行的干预策略。

关键词: 尿源干细胞, 运动训练, 原代培养, 取材时间, 细胞增殖, 干性维持

Abstract: OBJECTIVE：To evaluate the impact of exercise training on primary yield and early biological quality of urine-derived stem cells (USCs) from a single volunteer and to determine feasibility of collecting samples at a fixed evening interval (20:00-21:00)，thereby providing guidance for non-invasive and standardized USCs isolation. METHODS：A self-controlled design was adopted. Urine samples were collected from the same volunteer at four time points-early morning (6:00-7:00)，forenoon (9:00-10:00)，afternoon (14:00-15:00) and evening (20:00-21:00)-to compare primary clone-forming efficiency and identify the optimal sampling period. Within this period，the volunteer first maintained normal daily activity for 7 days (rest group) and then performed a standardized run (2.8 km in 16-18 min) daily for another 7 days (exercise group)，with urine collected immediately post-exercise. USCs were isolated and cultured under identical protocols. Cellular morphology was assessed by morphological observation. Cell proliferation was assessed by MTT assay over 7 days to prepare growth curves. RT-qPCR was employed to measure mRNA levels of the pluripotency genes OCT4，C-MYC，SOX2 and NANOG，while Western blot was used to evaluate OCT4 and C-MYC protein expression. RESULTS：Evening collection (20:00-21:00) consistently yielded USC clones with the highest formation rate. Within this window，exercise-derived USCs displayed homogeneous morphology and lower senescence versus sedentary controls. The proliferation curve revealed a significantly higher D(570) on day 7 (P<0.01)，indicating sustained proliferative capacity. OCT4 and C-MYC mRNA levels were up-regulated (P<0.05 and P<0.01，respectively)，whereas SOX2 and NANOG mRNA levels remained unchanged (P>0.05). Consistently，OCT4 and C-MYC protein expression were markedly elevated (P<0.01). CONCLUSION：Evening urine collection (20:00-21:00) combined with exercise training significantly improved USC extraction efficiency and enhanced their early biological characteristics，offering a simple and feasible strategy for subsequent scalable expansion and clinical translation.

Key words: urine-derived stem cells, exercise training, primary culture, sampling time, cell proliferation, stemness maintenance

中图分类号:

R329.2+8

徐嘉琪, 王凤龙, 孙震晓. 运动训练对尿源干细胞原代产量及早期生物学特性的影响[J]. 癌变·畸变·突变, 2025, 37(6): 473-477,482.

XU Jiaqi, WANG Fenglong, SUN Zhenxiao. Impact of exercise training on primary yield and biological quality of urine-derived stem cells[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(6): 473-477,482.

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