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溶血磷脂酸对离体培养的大鼠胚胎神经干细胞向神经胶质细胞分化的影响

崔慧林, 乔健天

山西医科大学神经生物学研究室.山西,太原 030001

摘要

该研究用免疫细胞化学荧光双标技术观察了溶血磷脂酸(lysophosphatidic acid,LPA)对大鼠胚胎神经干细胞(neural stem cells,NSCs)分化为少突胶质细胞(galactocerebroside-positive,Gal--C阳性)和星形胶质细胞(glial fibrillary acidic protein--positive,GFAP阳性)的影响,并且用RT--PCR技术对NSCs可能表达的LPA受体进行分析。结果显示:(1)加入不同浓度(0.01---3.0#mu#mol/ L)LPA,d7进行检测时,少突胶质细胞数量呈明显的剂量依赖性增加,峰值出现在1.0#mu#mol/L LPA组,少突胶质细胞所占百分比从对照组的8.5%增加到32.6%;(2)星形胶质细胞的分化几乎不受LPA的影响,第7天时各LPA处理级星形胶质细胞百分比与对照组相比均无显著性差异;(3)RT--PCR结果显示,大鼠胚胎NSCs的LPA_(1)和LPA_(3)受体表达明显,而LPA_(2)受体表达很弱。以上结果表明,较低浓度的LPA可能作为细胞外信号,通过LPA_(1)和LPA_(3)受体促进大鼠胚胎NSCs向少突胶质细胞分化和生成,但对星形胶质细胞的分化过程无明显影响。

关键词： 分化; 神经胶质细胞; 神经干细胞; 溶血磷脂酸; 溶血磷脂酸受体

Effect of lysophosphatidic acid on differentiation of embryonic neural stem cells into neuroglial cells in rats {sl in vitro}

Cui Huilin, Qiao Jiantian

Department of Neurobiology,Shanxi Medical University.Taiyuan 030001,Shanxi

Abstract

To study the effect of lysophosphatidic acid (LPA) on the differentiation of embryonic neural stem cells (NSCs) into neuroglia cells in rats in vitro, both oligodendrocytes and astrocytes were detected by their marker proteins galactocerebroside (Gal-C) and glia fibrillary acidic protein (GFAP), respectively, using double-labeling immunocytochemistry. RT-PCR assay was also used for analyzing the expression of LPA receptors in NSCs. Our results showed that: (1) Different concentrations of LPA (0.05-3.0 #mu#mol/L) were added to culture media and cell counting was carried out on the 7th day in all groups. Exposure to LPA led to a dose-dependent increase of oligodendrocytes with the response peaked at 1.0 #mu#mol/L, with an increased percentage of 32.6% (P<0.01) of total cells as compared to that of 8.5% in the vehicle group; (2) LPA showed no effect on the differentiation of NSCs into astrocytes; (3) RT-PCR assay showed that LPA_(1) and LPA_(3) receptors were strongly expressed while LPA_(2) receptor expressed weakly in NSCs. These results suggest that LPA at low concentration might act as an extracellular signal through the receptors in NSCs, mainly LPA_(1) and LPA_(3) receptors, to promote the differentiation of NSCs into oligodendrocytes, while it exhibits little, if any, conceivable effect on the differentiation of NSCs into astrocytes.

Key words: Differentiation;neuroglial cells;neural stem cells;Lysophosphatidic acid;lysophosphatidic acid receptors

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引用本文：

崔慧林, 乔健天. 溶血磷脂酸对离体培养的大鼠胚胎神经干细胞向神经胶质细胞分化的影响[J]. 生理学报 2007; 59 (6): 759-764.

Cui Huilin, Qiao Jiantian. Effect of lysophosphatidic acid on differentiation of embryonic neural stem cells into neuroglial cells in rats {sl in vitro}. Acta Physiol Sin 2007; 59 (6): 759-764 (in Chinese with English abstract).