黄精降低活性氧水平促进衰老内皮祖细胞功能的研究
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摘要
目的观察黄精在老年大鼠内皮祖细胞(endothelial progenitor cells,EPCs)传代培养衰老进程中对细胞功能及活性氧(reactive oxygen species,ROS)水平的影响。方法分离培养大鼠骨髓EPCs并鉴定,将培养至第2代的EPCs分为4组,分别用黄精含药血清和空白对照血清继续培养至第4、6、8代。β-半乳糖苷酶染色法检测细胞衰老程度;MTT法检测细胞增殖功能;Transwell小室检测迁移功能;体外成血管试剂盒检测成血管功能;流式细胞仪检测ROS水平。结果 EPCs在体外传代培养中,其衰老程度逐渐加重,并伴有增殖、迁移和成血管功能的明显减退,ROS水平急剧升高(P<0.05);经黄精干预后,EPCs衰老程度有所减轻,其功能有所增强,ROS水平明显降低(P<0.05)。结论黄精可能通过降低ROS水平来延缓传代培养EPCs的衰老进程,保护EPCs的功能。
Aim To investigate the effect of Rhizoma Polygonati(RP) on the functional activity and the level of reactive oxygen species(ROS) on the subculture of endothelial progenitor cells(EPCs) in aging rats. Methods EPCs from bone marrow in rats were isolated and cultured in vitro and identified. The 2 nd EPCs were divided into fourgroups, which were subcultured to the 4 th, 6 th and 8 th passage with application of drug-serum containing RP or not. The positive rate of cell senescence was detected by beta-galactosidase, the proliferation, migration and tubule formation were assayed by MTT, transwell chamber and in vivo angiogenesis kit, respectively, and ROS level was detected by flow cytometry. Results In the process of EPCs subculture, the positive rate of cell senescence gradually increased, accompanied by a significant decrease in proliferation, migration and tubule formation. The level of ROS increased sharply(P<0.05). RP could reduce the positive rate of cell senescence, enhance the functional activities, and decrease the level of ROS of EPCs significantly(P<0.05). Conclusions RP may delay the aging process of EPCs in vitro and protect the function of EPCs by decreasing the level of ROS.
Aim To investigate the effect of Rhizoma Polygonati(RP) on the functional activity and the level of reactive oxygen species(ROS) on the subculture of endothelial progenitor cells(EPCs) in aging rats. Methods EPCs from bone marrow in rats were isolated and cultured in vitro and identified. The 2 nd EPCs were divided into fourgroups, which were subcultured to the 4 th, 6 th and 8 th passage with application of drug-serum containing RP or not. The positive rate of cell senescence was detected by beta-galactosidase, the proliferation, migration and tubule formation were assayed by MTT, transwell chamber and in vivo angiogenesis kit, respectively, and ROS level was detected by flow cytometry. Results In the process of EPCs subculture, the positive rate of cell senescence gradually increased, accompanied by a significant decrease in proliferation, migration and tubule formation. The level of ROS increased sharply(P<0.05). RP could reduce the positive rate of cell senescence, enhance the functional activities, and decrease the level of ROS of EPCs significantly(P<0.05). Conclusions RP may delay the aging process of EPCs in vitro and protect the function of EPCs by decreasing the level of ROS.
引文
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[10] 雷升萍,王靓,龙子江,等.黄精多糖通过TLR4-MyD88-NF-κB通路抑制缺氧/复氧H9c2心肌细胞炎性因子释放[J]. 中国药理学通报,2017,33(2):255-60.
[10] Lei S P,Wang L,Long Z J,et al. Inhibitory effect of Polygonatum sibiricum polysaccharides on release of inflammatory cytokines of anoxia/reoxygenation H9c2 myocardial cells through TLR4-MyD88-NF-κB signaling pathway[J]. Chin Pharmacol Bull,2017,33(2):225-60.
[2] Assmus B,Honold J,Schachinger V,et al. Transcoronary transplantation of progenitor cells after myocardial infarction[J].N Engl J Med,2006,355(12):1222-32.
[3] 宰青青,秦臻,叶兰.黄精对自然衰老大鼠内皮祖细胞功能及端粒酶活性的影响[J].中国中西医结合杂志,2016,36(12):1480-5.
[3] Zai Q Q,Qin Z,Ye L. Effect of Rhiaoma Polygonati on the function of endothelial progenitor cells and telomerase activity in nature senescent rats[J].Chin J Integr Tradit West Med,2016,36(12):1480-5.
[4] 秦臻,黄水清.当归补血汤含药血清对内皮祖细胞功能及其PI3K/Akt通路影响的研究[J].中国药理学通报,2013,29(9):1320-3.
[4] Qin Z,Huang S Q. Influence of drug-serum containing Dangguibuxue decoction on fuctional activity and PI3K/Akt pathway of endothelial progenitor cells[J].Chin Pharmacol Bull,2013,29(9):1320-3.
[5] Zampetaki A,Kirton J P,Xu Q. Vascular repair by endothelial progenitor cells[J].Cardiovasc Res,2008,78(3):413-21.
[6] Umemura T,Soga J,Hidaka T,et al. Aging and hypertension are independent risk factors for reduced number of circulating endothelial progenitor cells[J].Am J Hypertens,2008,21(11):1203-9.
[7] Imanishi T,Tsujioka H,Akasaka T. Endothelial progenitor cells dysfunction and senescence: contribution to oxidative stress[J].Curr Cardiol Rev,2008,4(4):275-86.
[8] 陈永红,杜冠华.线粒体与衰老[J].中国药理学通报,2000,16(5):485-8.
[8] Chen Y H,Du G H. Mitochondria and aging[J].Chin Pharmacol Bull,2000,16(5):485-8.
[9] 马志茹,袁倬斌. 电化学法研究黄精及枸杞子对活性氧自由基的清除作用[J].中国药学杂志,1999,34(10):665-7.
[9] Ma Z R,Yuan Z B. Scavenging effect of fructus lycii and rhizoma polygonati on active oxygen free radicals[J].Chin Pharmacol J,1999,34(10):665-7.
[10] 雷升萍,王靓,龙子江,等.黄精多糖通过TLR4-MyD88-NF-κB通路抑制缺氧/复氧H9c2心肌细胞炎性因子释放[J]. 中国药理学通报,2017,33(2):255-60.
[10] Lei S P,Wang L,Long Z J,et al. Inhibitory effect of Polygonatum sibiricum polysaccharides on release of inflammatory cytokines of anoxia/reoxygenation H9c2 myocardial cells through TLR4-MyD88-NF-κB signaling pathway[J]. Chin Pharmacol Bull,2017,33(2):225-60.