生理学报
生理學報
생이학보
ACTA PHYSIOLOGICA SINICA
2007年
6期
759-764
,共6页
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
分化%神經膠質細胞%神經榦細胞%溶血燐脂痠%溶血燐脂痠受體
분화%신경효질세포%신경간세포%용혈린지산%용혈린지산수체
differentiation%neuroglial cells%neural stem cells%lysophosphatidic acid%lysophosphatidic acid receptors
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处理组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LPA1和LPA3受体表达明显,而LPA2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LPA1和LPA3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.
本研究用免疫細胞化學熒光雙標技術觀察瞭溶血燐脂痠(lysophosphatidic acid,LPA)對大鼠胚胎神經榦細胞(neural stem cells,NSCs)分化為少突膠質細胞(galactocerebroside-positive,Gal-C暘性)和星形膠質細胞(glial fibrillary acidic protein-positive,GFAP暘性)的影響,併且用RT-PCR技術對NSCs可能錶達的LPA受體進行分析.結果顯示:(1)加入不同濃度(0.01~3.0μmol/L)LPA,第7天進行檢測時,少突膠質細胞數量呈明顯的劑量依賴性增加,峰值齣現在1.0μmol/L LPA組,少突膠質細胞所佔百分比從對照組的8.5%增加到32.6%:(2)星形膠質細胞的分化幾乎不受LPA的影響,第7天時各LPA處理組星形膠質細胞百分比與對照組相比均無顯著性差異;(3)RT-PCR結果顯示,大鼠胚胎NSCs的LPA1和LPA3受體錶達明顯,而LPA2受體錶達很弱.以上結果錶明,較低濃度的LPA可能作為細胞外信號,通過LPA1和LPA3受體促進大鼠胚胎NSCs嚮少突膠質細胞分化和生成,但對星形膠質細胞的分化過程無明顯影響.
본연구용면역세포화학형광쌍표기술관찰료용혈린지산(lysophosphatidic acid,LPA)대대서배태신경간세포(neural stem cells,NSCs)분화위소돌효질세포(galactocerebroside-positive,Gal-C양성)화성형효질세포(glial fibrillary acidic protein-positive,GFAP양성)적영향,병차용RT-PCR기술대NSCs가능표체적LPA수체진행분석.결과현시:(1)가입불동농도(0.01~3.0μmol/L)LPA,제7천진행검측시,소돌효질세포수량정명현적제량의뢰성증가,봉치출현재1.0μmol/L LPA조,소돌효질세포소점백분비종대조조적8.5%증가도32.6%:(2)성형효질세포적분화궤호불수LPA적영향,제7천시각LPA처리조성형효질세포백분비여대조조상비균무현저성차이;(3)RT-PCR결과현시,대서배태NSCs적LPA1화LPA3수체표체명현,이LPA2수체표체흔약.이상결과표명,교저농도적LPA가능작위세포외신호,통과LPA1화LPA3수체촉진대서배태NSCs향소돌효질세포분화화생성,단대성형효질세포적분화과정무명현영향.
To study the effect of lysophosphatidic acid(LPA) on the differentiation of embryonic neural stem cells(NSCs)into neuroglial cells in rats in vitro,both oligodendrocytes and astrocytes were detected by their marker proteins galactocerebroside(GalC)and glial 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)LPA at different concentrations(0.01-3.0 μmol/L)was added to culture medium 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 μmol/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 LPA1 and LPA3 receptors were strongly expressed while LPA2 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 LPA1 and LPA3 receptors,to promote the differentiation of NSCs into oligodendrocytes,while it exhibits little,if any,conceivable effect on the differentiation of NSCs into astrocytes.
