摘要
目的观察Notch-1对嗅球成鞘细胞(olfactory ensheathing cells,OECs)神经生长因子(nerve growth factor,NGF)表达的影响。方法原代培养大鼠OECs,用有活性的Notch质粒(NotchICV),无活性的Notch质粒(NotchVK)以及空质粒分别转染OECs,将实验对象分成NotchICV转染组,NotchVK转染组和空白质粒组,应用免疫荧光双标、Western blot及RT-PCR技术,检测各组OECs中NGF及其mRNA的表达变化。结果转染Notch-1后,OECs中NGF的表达明显增高。免疫荧光双标结果显示,NotchICV转染组OECs中NGF的表达较NotchVK转染组和空白质粒组均明显增高(P<0.05);RT-PCR结果显示,NotchICV转染组OECs中NGF mRNA的表达(0.4961±0.0269)均高于NotchVK转染组(0.2445±0.0134)及空白质粒组(0.2459±0.0160)(P<0.01);Western blot结果显示,NotchICV转染组OECs中NGF的表达(0.8875±0.0166)均高于NotchVK转染组(0.6862±0.0592)和空白质粒组(0.6821±0.0670)(P<0.01)。结论 Notch-1转染OECs后,其NGF的表达上调,提示Notch-1能增强OECs中NGF的合成。
Objective To observe the effect of Notch-1 on expression of nerve growth factor（NGF）in olfactory ensheathing cells（OEC）of rats.Methods Rats were divided into active Notch plasmid（NotchICV）group,inactive Notch plasmid（NotchVK）group,and empty plasmid group.NotchICV,NotchVK,and empty plasmid were transfected into OEC of rats.Immunofluorescence,Western blotting and RT-PCR were used respectively to detect the expression of NGF and NGF mRNA in OEC of rats.Results Immunofluorescence showed that the expression level of NGF was significantly higher in NotchICV group than in NotchVK group and empty plasmid group（P0.05）.RT-PCR demonstrated that the expression level of NGF mRNA was significantly higher in NotchICV group（0.496 1±0.026 9）than in NotchVK group（0.244 5±0.013 4）and empty plasmid group（0.245 9±0.016 0）（P0.01）.Western blotting displayed that the expression level of NGF was significantly higher in NotchICV group（0.887 5±0.016 6）than in NotchVK group（0.686 2±0.059 2）and empty plasmid group（0.682 1±0.067 0）（P0.01）.Conclusion Expression of NGF is up-regulated in Notch-1 transfected OEC,and Notch-1 can increase the synthesis of NGF in OEC.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2010年第21期2290-2293,共4页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(30670743)
重庆市自然科学基金(2008BB5224)~~
