溶血磷脂酸对嗅鞘细胞形态增殖及表达脑源性神经营养因子的影响

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1、溶血磷脂酸对嗅鞘细胞形态增殖及表达脑源性神经营养因子的影响【摘要】观察溶血磷脂酸(lysophosphatidicacid，LPA)对体外培养的嗅鞘细胞(olfactoryensheathingcells，OECs)形态、增殖和表达脑源性神经营养因子(brainderivedneurotropicfactor，BDNF)的影响。［方法］取成年大鼠原代OECs经纯化后培养，分别用1、5、10、20、50μm／L的LPA干预一定时间后，用免疫荧光染色法、MTT、WesternBlot技术分别观察OECs形态变化、增殖和表达BDNF的情况。［结果］LPA使O

2、ECs形态由突起形向扁平形转变，去除LPA可逆转这种转变；1～50μm／L的LPA均能促进其增殖，干预后60h达到增殖高峰，浓度为10μm／L时促增殖作用最显著;1～50μm／L的LPA均可上调其表达BDNF。［结论］LPA能使OECs形态由突起形向扁平形发生可逆性变化；能促进OECs增殖并具有时间和浓度依赖性；能上调OECs表达BDNF。【关键词】溶血磷脂酸嗅鞘细胞Abstract：［Objective］Toinvestigatetheeffectsoflysophosphatidicacid(LPA)onthemorphology,prolifera

3、tionandbrainderivedneurotropicfactor(BDNF)expressionofolfactoryensheathingcells(OECs)invitro.［Method］PrimaryculturesofOECsseparatedfromadultratolfactorybulbswere16purifiedandcultured.FiveexperimentalculturesweregrownforaperiodoftimeinmediawithLPAatdifferentconcentrations,namely1

4、,5,10,20and50μmol/L,andthecontrolculturewasgrowninthemediumwithoutLPA.ImmunofluorescentstainingwasusedtoidentifyOECsandtoobservetheirmorphologicalchanges.TheproliferationofOECswasmeasuredbyMTTassay.WesternblottingwasusedtodetecttheproteinexpressionofBDNF.［Result］ExposuretoLPAinme

5、diuminducedtheswitchinthedominantmorphologyofOECsfromprocessbearingtoflatmorphology.ThisshiftinmorphologywasreversedwhenLPAwasremovedfrommedia.LPAatconcentrationsfrom1μmol/Lto50μmol/LenhancedOECsproliferation,especiallyattheconcentrationof10μm/L.ProliferationofOECsinallexperimen

6、talculturesreachedtheirrespectivesignificantpeaksafter60hofLPAtreatment.ThereweresignificantupregulationsinBDNFexpressionofOECstreatedwithLPA(1～50μm/L)comparedwiththoseinthecontrolculture.［Conclusion］Areversiblechangefromprocess-bearingtoflatinmorphologyofOECscanbeinducedbyLPA.LP

7、AstimulatesOECsproliferationinatimeandconcentrationdependentmanner.LPAupregulatesBDNFexpressionofOECs.16Keywords：lysophosphatidicacid;olfactoryensheathingcells;morphology;proliferation;brain-derivedneurotropicfactor尽管OECs用于中枢神经损伤修复研究己近20年，并被普遍认为是最有应用前景的候选细胞［1］，然而对其某些基本的生物学特性如免疫

8、学与形态学关系的认识仍然存在分歧［2，3］。此外，OECs神经修复作用受到其表达