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本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(0.01～3.0μmol/ L)LPA,第7天进行检测时,少突胶质细胞数量呈明显的剂量依赖性增加,峰值出现在1.0μmol/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。
In this study, the effects of lysophosphatidic acid (LPA) on the differentiation of rat embryonic neural stem cells (NSCs) into oligodendrocytes (galactocerebroside-positive, Gal-C positive) ) And glial fibrillary acidic protein-positive (GFAP-positive), and the LPA receptors that may be expressed by NSCs were analyzed by RT-PCR. The results showed that: (1) LPA at different concentration (0.01-3.0μmol / L) was detected in the seventh day, and the number of oligodendrocytes increased in a dose-dependent manner. The peak appeared in 1.0μmol / L LPA group, The percentage of oligodendrocytes increased from 8.5% to 32.6% in the control group. (2) The differentiation of astrocytes was hardly affected by LPA. The percentage of LPA-treated astrocytes on day 7 was (3) The results of RT-PCR showed that the expression of LPA_1 and LPA_3 receptors in rat embryo NSCs was obvious, while the expression of LPA_2 receptors was weak. These results indicated that LPA at a lower concentration may act as an extracellular signal to promote the differentiation and generation of NSCs into oligodendrocytes by LPA_1 and LPA_3 receptors but not to the differentiation of astrocytes.