中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5992-5996.doi: 10.3969/j.issn.1673-8225.2010.32.022

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

残粒脂蛋白诱导大鼠脂肪间充质干细胞的成脂分化

马美芳，张  茜，刘  玲   

  1. 中南大学湘雅二医院心内科，湖南省长沙市 410011
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 通讯作者: 刘玲，博士，副主任医师，中南大学湘雅二医院心内科，湖南省长沙市 410011 feliuling@ medmail.com.cn
  • 作者简介:马美芳★，女，1981年生，河北省邯郸市人，汉族，2010年中南大学毕业，硕士，医师，主要从事残粒脂蛋白致动脉粥样硬化机制方面的研究。 wish0818@163. com
  • 基金资助:

    中华医学会动脉粥样硬化研究专项资金课题（09010570212），课题名称“残粒脂蛋白对脂肪间充质干细胞成脂分化的影响以及阿托伐他汀的干预作用”；中南大学湘雅二医院代谢综合征研究中心资助课题（DY-2008-01-01），课题名称“残粒脂蛋白对脂肪间充质干细胞成脂分化的影响”。

Remnant lipoprotein induces adipogenic differentiation of rat adipose tissue-derived mesenchymal stem cells

Ma Mei-fang, Zhang Qian, Liu Ling   

  1. Department of Cardiology, Second Xiangya Hospital, Central South University, Changsha  410011, Hunan Province, China
  • Online:2010-08-06 Published:2010-08-06
  • Contact: Liu Ling, Doctor, Associate chief physician, Department of Cardiology, Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China feliuling@medmail. com.cn
  • About author:Ma Mei-fang★, Master, Physician, Department of Cardiology, Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China Department of wish0818@163.com
  • Supported by:

     the Atherosclerosis Research Special-Purpose Foundation of Chinese Medical Association, No. 09010570212; the grant from the Metabolic Syndrome  Research Center of Second Xiangya Hospital of Central South University, No. DY-2008-01-01

PDF

349

被引次数

5

摘要：

背景：高三酰甘油血症患者血浆中的极低密度脂蛋白和乳糜微粒增多，可被水解为残粒脂蛋白。极低密度脂蛋白可诱导其他前体脂肪细胞的成脂分化，推测残粒脂蛋白也具有诱导脂肪间充质干细胞成脂分化的能力。
目的：探讨不同质量浓度残粒脂蛋白对大鼠脂肪间充质干细胞成脂分化的影响。
方法：无菌条件下切取大鼠腹股沟脂肪垫，胶原酶消化法获取脂肪间充质干细胞，接种入含胎牛血清的DMEM/F12培养基进行培养，观察细胞形态变化，流式细胞仪检测细胞表面分子的表达。将传至第2代的脂肪间充质干细胞分为4组，对照组单纯以10 mg/L胰岛素进行孵育，剩余3组另加入50，100，200 mg/L残粒脂蛋白，18 d后通过油红O染色评价成脂分化情况。
结果与结论：原代脂肪间充质干细胞早期呈集落样生长，随传代次数增加，逐渐呈均一性梭状细胞外观，细胞表达CD105，基本不表达CD34。与对照组比较，加入50 mg/L残粒脂蛋白后油红O染色强度无明显变化(P > 0.05)，加入100 mg/L残粒脂蛋白后油红O染色强度明显升高(P < 0.05)；加入200 mg/L残粒脂蛋白油红O染色强度明显高于其他3组(P < 0.05)。提示残粒脂蛋白以浓度依赖性的方式诱导脂肪间充质干细胞成脂分化。

关键词: 残粒脂蛋白, 成脂分化, 胰岛素, 脂肪间充质干细胞, 干细胞

Abstract:

BACKGROUND:Very low density lipoprotein and chylomicron are increased in blood plasma of patients with hypertriacylglycerolemia, and can be hydrolyzed into remnant lipoprotein. Very low density lipoprotein can induce the adipogenic differentiation of other preadipocytes. It is presumed that remnant lipoprotein also can induce the adipogenic differentiation of adipose tissue-derived mesenchymal stem cells (ADMSCs).
OBJECTIVE:To explore the effects of different concentrations of remnant lipoprotein on the adipogenic differentiation of ADMSCs.
METHODS: Collagenase type II was used to isolate ADMSCs from inguinal adipose tissue of rats under sterile condition. The cells were cultured in the DMEM/F12 medium supplemented with fetal bovine serum. The morphological changes of ADMSCs were observed. Flow cytometry was performed to determine cell surface molecules. The second passage of ADMSCs was divided into four groups. Cells in the control group were incubated in 10 mg/L insulin. 50, 100, 200 mg/L remnant lipoprotein was added in the other three groups. Lipid droplets within cells were identified by Oil red O staining 18 days later.
RESULTS AND CONCLUSION:  Primary ADMSCs grew in a manner like colony. With the increased passage, ADMSCs looked like spindle fibroblasts and were positive for CD105 but negative for CD34. Compared with the control group, no significant difference in the strength of Oil red O staining was detected following an addition of 50 mg/L remnant lipoprotein (P > 0.05), and the strength of Oil red O staining was significantly increased following an additional 100 mg/L remnant lipoprotein (P < 0.05). The strength was significantly greater in 200 mg/L remnant lipoprotein group than other three groups (P < 0.05). Results have suggested that remnant lipoprotein induces adipogenic differentiation of ADMSCs in a concentration dependent manner.

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