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纸质出版:2013
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张蔓,舒秀兰,李文迪,刘晶晶,陈洁,刘友学.atRA抑制大鼠骨髓间充质干细胞成脂分化[J].陆军军医大学学报(原第三军医大学学报),2013,35(15):1574-1578.
Zhang Man,Shu Xiulan,Li Wendi,Liu Jingjing,Chen Jie,Liu Youxue.AtRA inhibits adipogenic differentiation of bone marrow mesenchymal stem cells in rats[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(15):1574-1578.
张蔓,舒秀兰,李文迪,刘晶晶,陈洁,刘友学.atRA抑制大鼠骨髓间充质干细胞成脂分化[J].陆军军医大学学报(原第三军医大学学报),2013,35(15):1574-1578. DOI:
Zhang Man,Shu Xiulan,Li Wendi,Liu Jingjing,Chen Jie,Liu Youxue.AtRA inhibits adipogenic differentiation of bone marrow mesenchymal stem cells in rats[J].J Amry Med Univ (J Third Mil Med Univ),2013,35(15):1574-1578. DOI:
目的 探讨高浓度全反式视黄酸(all-trans retinoic acid
atRA)抑制大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells
BMSCs)向脂肪细胞分化的机制。 方法 体外分离、培养和诱导大鼠BMSCs;在成脂诱导液中加入0.1、0.5、1.0、5.0 μmol/L atRA,光学显微镜下观察细胞形态及油红O染色变化;油红O染色提取法分析不同浓度atRA对BMSCs成脂分化的影响;real-time PCR测定细胞视黄酸受体(RARα、RARγ)、CCAAT增强子结合蛋白α(CEBPα)及过氧化物酶体增殖激活受体γ2(PPARγ2)mRNA水平的变化。 结果 细胞形态及油红O染色观察发现,在0.1、0.5、1.0、5.0 μmol/L atRA作用下大鼠BMSCs向脂肪细胞分化受到抑制,且呈浓度依赖性;油红O染色提取法半定量测定结果显示,随着atRA浓度升高,分化细胞内脂肪含量降低(
P
0.01);real-time PCR检测显示与血清组(未加atRA)相比,加入atRA后,大鼠BMSCs的CEBPα和PPARγ2表达显著降低(
P
0.05),RARα表达无显著差异[血清组vs 1.0 μmol/L atRA组: (0.008 4±0.001 9) vs (0.009 1±0.001 3),
P
0.05
]
,但RARγ表达显著增加[血清组vs 1.0 μmol/L atRA组: (0.022 6±0.001 2) vs (0.053 1±0.002 3),
P
0.01
]
。 结论 0.1、0.5、1.0、5.0 μmol/L atRA可抑制大鼠BMSCs向脂肪细胞分化,其机制可能与RARγ的上调及CEBPα、PPARγ2的下调有关。
Objective To investigate the mechanism of all-trans retinoic acid (atRA) to inhibit the adipogenic differentiation of rat bone marrow mesenchymal stem cells (BMSCs). Methods Rat BMSCs were isolated
incubated and induced to differentiate into adipocytes. AtRA at high concen
tration (0.1-5.0 μmol/L) was added to the medium for adipogenic induction. The morphologic changes of BMSCs during differentiation were observed under a light microscope. The effect of atRA on the adipogenic differentiation was analyzed by oil red O staining. The expression of retinoic acid receptors α and γ (RARα and RARγ)
CCAAT/enhancer binding protein a (CEBPα) and peroxisome proliferators-activated receptor γ2 (PPARγ2) was analyzed by real-time PCR. Results At the concentration of 0.1-5.0 μmol/L
atRA inhibited rBMSCs from differentiating into adipocytes in a dose-dependent manner. The fat content in differentiated cells was decreased along with the increase of atRA concentration (
P
0.05). Compared with the control group (without adding atRA)
the mRNA expression levels of CEBPα and PPARγ2 were decreased significantly in the atRA groups (
P
0.05). Compared with the control group
the mRNA expression of RARα in the atRA group (1.0 μmol/L) was not significantly different (0.008 4±0.001 9 vs 0.009 1±0.001 3
P
0.05)
while the mRNA expression of RARγ significantly was increased (0.022 6±0.001 2 vs 0.053 1±0.002 3
P
0.01). Conclusion AtRA (0.1-5.0 μmol/L) can inhibit the adipogenic differentiation of rBMSCs by up-regulating the expression of RARγ and down-regulating the expression of CEBPα and PPARγ2.
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