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目的：动态观察大鼠自然衰老进程中骨髓内皮祖细胞（endothelial progenitor cells,EPCs）衰老程度、细胞功能、线粒体结构和功能的变化及黄精的干预效果。方法：将96只18月龄的大鼠按体质量随机分为老年组、黄精低剂量（1 g/kg）、中剂量（2 g/kg）、高剂量组（4 g/kg），每组24只；另设24只2月龄大鼠为青年组。各组每天灌胃1次，持续12周，在第4周、8周、12周每组各取8只大鼠麻醉处死，分离培养骨髓EPCs并鉴定；采用β-半乳糖苷酶染色法检测细胞衰老程度，采用CCK-8法、Transwell小室、体外成血管试剂盒检测细胞的增殖、迁移和成小管功能；流式细胞仪检测胞内活性氧（reactive oxygen species,ROS）水平；JC-1法检测线粒体膜电位（mitochondrial membrane potential,MMP）；化学发光法检测线粒体ATP含量；透射电镜观察线粒体形态结构。结果：随周龄增加，老年组大鼠骨髓EPCs衰老程度逐渐加重，细胞增殖、迁移和成小管功能明显减退；胞内ROS水平升高且ATP含量及MMP降低（P<0.05）；电镜下可见线粒体固缩和大量自噬体。经黄精干预后，老年大鼠骨髓EPCs衰老程度减轻，细胞增殖、迁移和成小管功能有所改善；胞内ROS水平降低且ATP含量及MMP上升（P<0.05）；电镜下线粒体形态结构较为正常，可见少量自噬体。结论：黄精可通过维护线粒体结构和功能的完整性来促进衰老大鼠骨髓EPCs的功能。
Abstract：Objective: To dynamically observe the effect of Rhizoma Polygonati(RP) on the senescence degree, the functional activities, the mitochondrial function and structure of endothelial progenitor cells(EPCs) derived from bone marrow in the natural aging process of rats. The intervention effect of RP was also observed. Methods: A total of 96 rats(18 months) were randomly divided into four groups: an old group, a RP-low-dose group(1 g/kg), a RP-mid-dose group(2 g/kg), and a RP-high-dose group(4 g/kg), another 24 rats(2 months) were used as the young group. All rats were administered by gavage once a day for 12weeks. At the 4th, 8th, 12th week, 8 rats in each group were anesthetized and euthanized, then EPCs were isolated, cultured and indentified. EPCs senescence was assessed by beta-galactosidase, cellular proliferation, migration, tubule formation were evaluated by CCK-8,transwell chamber, and in vivo angiogenesis kit, respectively. The level of reactive oxygen species(ROS) was assessed by flow cytometry, the mitochondrial membrane potential(MMP) was measured by JC-1, and the ATP content was evaluated by chemiluminescence; the structure of mitochondria was observed by transmission electron microscopy. Results: In the old group, EPCs senescence increases with increasing week age, accompanied by a significant decrease in proliferation, migration and tubule formation. The level of ROS increased but the ATP content and MMP both decreased(P<0.05). The mitochondria was solidified and there were more autophagosomes. RP treatment could reduce the positive rate of cell senescence, promote the functional activities of EPCs, decrease the ROS level, and increase the ATP content and MMP(P<0.05). The mitochondria structure became regular, and a few autophagosomes were observed. Conclusion: RP could promote the functional activities of EPCs in the natural ageing process of rats by maintaining the integrity of mitochondrial structure and restoration of mitochondrial function.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20240904.001
中图分类号:R285.5
引用信息:
[1]秦臻,叶利兵,冯静月,等.黄精维护线粒体结构功能促进衰老大鼠骨髓内皮祖细胞功能的研究[J].海南医科大学学报,2025,31(07):497-505+512.DOI:10.13210/j.cnki.jhmu.20240904.001.
基金信息:
国家自然科学基金资助项目(81860872,82360964); 贵州省中医药管理局中医药、民族医药科学技术研究课题(QZYY-2024-012)~~
2024-09-04
2024-09-04
2024-09-04