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|本期目录/Table of Contents|

丹参素促大鼠心肌细胞及心脏干细胞增殖的作用

《心脏杂志》[ISSN:1009-7236/CN:61-1268/R]

期数:
2013年第6期
页码:
644-648
栏目:
基础研究
出版日期:
2013-11-25

文章信息/Info

Title:
Effect of Danshensu on growth of myocardial cells and cardiac stem cells
作者:
宋 凡1; 2; 李岳洋1; 2; 孙纪元1; 2; 王四旺1; 2
(1.第四军医大学药学系天然药物学教研室；2.秦巴山区中药发展协同创新中心，陕西 西安 710032)
Author(s):
SONG Fan1; 2;  LI Yueyang1; 2;  SUN Ji yuan1; 2;  WANG Siwang1; 2
(1.Institute of Materia Medica, School of Pharmacy, Fourth Military Medical University, Xi’an 710032, Shaanxi, China; 2.Collaborative Innovation Center for Chinese Medicine in Qinba Mountains, Xi’an 710032, Shaanxi, China)
关键词:
心肌细胞; 心脏干细胞; 丹参素; 细胞增殖; 大鼠
Keywords:
myocardial cells;  cardiac stem cells;  Danshensu;  cell proliferation;  rat
分类号:
R284.1
DOI:
-
文献标识码:
A
摘要:
目的:大鼠原代心肌细胞及心脏干细胞（CSCs）培养条件的筛选及丹参素（DS-182）对两种细胞体外增殖的影响。方法:对比组织块培养法及差速培养法，选取适宜培养基、消化液浓度、消化时间、差速贴壁时间及心脏成纤维细胞（CFs）抑制剂等因素。分别用MTT比色法、ELISA法及流式细胞术(FCM)检测丹参素对大鼠心肌细胞及CSCs体外增殖的影响。结果:确定DMEM/F12 1∶1培养基、100 ml/L胎牛血清(FBS)、2.5 g/L胰蛋白酶、5 min/次×5次的消化时间及120 min的差速贴壁时间为最适分离纯化条件。MTT比色法检测显示，与空白对照组相比，经浓度为7.8×10-4~1×10-1 mol/L的DS-182处理12、24、48和72 h后，细胞的吸光度(A)值明显增加（P<0.05）。ELISA法检测显示，与空白对照组相比，经浓度为3.9×10-4~1×10-1 mol/L的DS182处理24 h后，细胞的A值明显增加（P<0.05）。FCM检测显示，经浓度为0.1 mol/L的DS-182处理24 h后，c-kit+的细胞数占细胞总数的比例增加了27.0%。结论:改良了一套较为完整的大鼠心肌细胞及CSCs的分离纯化方法。DS-182在一定浓度下（7.8×10-4~1×10-1 mol/L）对大鼠心肌细胞及CSCs的增殖具有明显的促进作用。
Abstract:
AIM:To explore the appropriate culture conditions and method for separating myocardial cells and cardiac stem cells (CSCs) from Sprague Dawley neonatal rat hearts and to study the effect of Danshensu (DS182) on the proliferation of the two kinds of cells. METHODS: Different culture mediums, digestive enzyme concentrations, digestion times, differential adhesion times and inhibitors of fibroblasts were selected to optimize the separation and purification methods of rat myocardial cells and CSCs. MTT assay, ELISA, FCM and other methods were used to determine the effect of DS182 on cell proliferation of rat myocardial cells and CSCs. RESULTS: Separation and purification conditions of the two kinds of cells were DMEM/F12 medium 1∶1, 100 ml/L FBS, 25 g/L trypsin, digestion time of 5 min for five times and the differential time of 120 min. MTT assay results showed that compared with those in the blank group, cell A value increased significantly when cells were exposed to the DS182 concentration of 78×10-4-1×10-1 mol/L at 12, 24, 48, 72 h after administration (P<0.05). ELISA results showed that cell OD value increased significantly when cells were exposed to the DS182 concentration of 3.9×10-4-1×10-1 mol/L at 24 h after administration (P<0.05). FCM results showed that the number of ckit+ increased by 27.0% when cells were exposed to the DS182 concentration of 0.1 mol/L at 24 h after administration. CONCLUSION: We improved the separation and purification methods of myocardial cells and CSCs. DS182 at certain concentration (78 × 10-4-1×10-1 mol/L) obviously promotes cell proliferation of myocardial cells and CSCs.

参考文献/References

[1]Lam FF,Yeung JH,Chan KM,et al.Relaxant efects of danshen aqueous extract and its constituent danshensu on rat coronary artery are mediated by inhibition of calcium channels[J].Vascul Phammeol,2007,46(4):271-277.
[2]Yang RX,Wang SY,Yan FF,et al.Danshensu protects vascular endothelia in a rat model of hyperhomocysteinemia[J].Acta Pharmacol Sin,2010,31(10):1395-1400.
[3]Le XY,Chen CL,Ma L,et al.Effects of Danshensu on the inci dence of ischemiarepefusion induced arrhythmia in hypertrophy rat heart[J].Chin J Nat Med,2008,6(6):461.
[4]Tang YQ,Wang MH,Le XY,et al.Antioxidant and cardioprotec·tive effects of Danshensu(3-(3,4- dihydroxypheny1)-2-hydroxypropanoic acid from Salvia miltiorrhiza)on isoproterenolinduced myocardial hypertrophy in rats[J].Phytomedicine,2011,18(12):1024-1030.
[5]Kajstura J,Leri A,Finato N,et al.Myocyte proliferation in endstage cardiac failure in humans[J].Proc Natl Acad Sci USA,1998,95(15):8801-8805.
[6]Bearzi C,Rota M,Hosoda T,et al.Human cardiac stem cells[J]. Proc Natl Acad Sci U S A,2007,104(35):14068-14073.
[7]康振军,钟 理,宋治远.Sca-1+心肌干细胞的研究进展[J].心血管病,2009,30(6):1047-1050.
[8]Zhang J,Huang C,Wu P,et al.Differentiation induction of cardiac c-kit positive cells from rat heart into sinus nodelike cells by 5-azacytidine [J].Tissue and Cell,2011,43(2):67-74.
[9]姜 宏,施广飞.心肌干细胞培养和纯化的实验方法研究[J].现代生物医学进展,2009,9(1):33-35.
[10]姜 宏,施广飞. 5-氮胞苷诱导心肌组织中c-kit+干细胞分化的实验研究[J].中国临床药理学与治疗学,2008,13(9):996-1001.
[11]王东进,施广飞,姜 宏.中药诱导干细胞分化为心肌细胞的效应[J].中国组织工程研究与临床康复,2010,14(45):8503-8506.
[12]康振峻,钟 理,宋治远.Sca-1+心肌干细胞的研究进展[J].心血管病学进展.2009,30(6):1047-1050.
[13]曹京燕,程月新,姜敏辉,等.5-氮胞苷体外诱导骨髓间充质干细胞向心肌样细胞的分化[J].心脏杂志,2009,21(6):801-804.
[14]郎明建,郭 敏,曾球棠,等.差速贴壁量和应用5-BrdU对心肌细胞的纯度及活力的影响[J].心脏杂志,2008,20(5):566-569.
[15]李宝善,马厚勋.骨髓间充质干细胞向心肌细胞的分化的调控机制及中药诱导研究进展[J].中国中西医结合杂志,2011,31(1):133-138.
[16]Liu J,Wang Y,Du W,et al.Sca-1-positive cardiac stem cell migration in a cardiac infarction model[J].Inflammation,2013,36(3):738-749.
[17]Golden HB,Gollapudi D,Gerilechaogetu F,et al.Isolation of cardiac myocytes and fibroblasts from neonatal rat pups[J].Methods Mol Biol,2012,843:205-214.

备注/Memo

备注/Memo:
收稿日期：2013-05-06.通讯作者：王四旺，教授，主要从事中药学研究Email：wangsiw@fmmu.edu.cn 共同通讯作者：孙纪元，副教授，主要从事中药学研究 Email：sunjiy@fmmu.edu.cn 作者简介：宋凡，硕士生 Email：songfan_000@163.com
更新日期/Last Update: 2013-11-20