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作 者:朱仲庚[1] 吴小涛[2] 管学能[3] 韦继南[2] 徐小彬[1]
机构地区:[1]上海市松江区中心医院骨科,201600 [2]东南大学附属中大医院骨科,南京 [3]江苏省中西医结合医院神经内科,南京
出 处:《中国矫形外科杂志》2008年第12期935-939,共5页Orthopedic Journal of China
摘 要:[目的]观察溶血磷脂酸(lysophosphatidic acid,LPA)对体外培养的嗅鞘细胞(olfactory ensheathing cells,OECs)形态、增殖和表达脑源性神经营养因子(brain-derived neurotropic factor,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、Western Blot技术分别观察OECs形态变化、增殖和表达BDNF的情况。[结果]LPA使OECs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。[ Objective] To investigate the effects of lysophosphatidic acid (LPA) on the morphology, proliferation and brainderived neurotropic factor ( BDNF ) expression of olfactory ensheathing cells ( OECs ) in vitro. [ Method ] Primary cultures of OECs separated from adult rat olfactory bulbs were purified and cultured. Five experimental cultures were grown for a period of time in media with LPA at different concentrations, namely 1, 5, 10, 20 and 50 μmol/L, and the control culture was grown in the medium without LPA. Immunofluorescent staining was used to identify OECs and to observe their morphological changes. The proliferation of OECs was measured by MTT assay. Western blotting was used to detect the protein expression of BDNF. [ Result] Exposure to LPA in medium induced the switch in the dominant morphology of OECs from process-bearing to fiat morphology. This shift in morphology was reversed when LPA was removed from media. LPA at concentrations from 1 μmol/L to 50 μmol/L enhanced OECs proliferation, especially at the concentration of 10 μm/L. Proliferation of OECs in all experimental cultures reached their respective significant peaks after 60 h of LPA treatment. There were significant upregulations in BDNF expression of OECs treated with LPA ( 1 - 50 μm/L) compared with those in the control culture. [ Conclusion] A reversible change from process -bearing to fiat in morphology of OECs can be induced by LPA. LPA stimulates OECs proliferation in a time- and concentration-dependent manner. LPA upregulates BDNF expression of OECs.
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