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5-脂氧酶/绿荧光蛋白转染评价PC12细胞损伤后5-脂氧酶核膜移位
  • ISSN号:1008-9292
  • 期刊名称:《浙江大学学报:医学版》
  • 时间:0
  • 分类:Q256[生物学—细胞生物学] Q421[生物学—神经生物学;生物学—生理学]
  • 作者机构:[1]浙江大学医学院药理学教研室,浙江杭州310058
  • 相关基金:国家自然科学基金(30371637)资助项目.
中文摘要:

目的:通过绿荧光蛋白(green fluorescence protein,GFP)/5-脂氧酶(5-lipoxygenase,5-LOX)稳定转染PC12细胞,以可视化方法观察5-LOX在不同损伤后的变化。方法:将带有5-LOX基因的pEGFP—C2质粒及pEGFP—C2质粒(对照),稳定转染至PC12细胞;在缺糖缺氧(oxygen—glucose deprivation,OGD)、H2O2和NMDA处理后,荧光显微镜下观察GFP/5-LOX在细胞内的定位;同时,以免疫细胞化学方法观察野生型5-LOX分布及其变化。结果:转染GFP/5-LOX的细胞内,GFP/5-LOX主要均匀表达于细胞核内;仅转染GFP的细胞GFP表达于胞核和胞浆。OGD和H2O2处理后,分别有50.6%和57.7%细胞的GFP/5-LOX移位到核膜;NMDA处理或仅转染GFP的细胞,未见核膜移位现象。野生型5-LOX分布在细胞核和细胞浆内,3种损伤处理均诱导核膜移位。结论:稳定转染GFP/5-LOX的PC12细胞,GFP/5-LOX主要分布在细胞核;不同损伤处理诱导的5-LOX核膜移位,有不同的特点。

英文摘要:

Objective: To evaluate the translocation of 5-lipoxygenase (5-LOX) after injuries by transfection with green fluorescence protein (GFP)/5-LOX in PC12 cells. Methods: PC12 cells were stably transfected with pEGFP-C2/5-LOX (GFP/5-LOX) or pEGFP-C2 vectors (control). After treatment with oxygen-glucose deprivation (OGD),H2O2 or NMDA,GFP/5-LOX localization in the cells was observed under a fluorescence microscope. Wild-type 5-LOX was determined by immunostaining after the treatment. Results. In the GFP/5-LOX-transfected cells, GFP/5-LOX was primarily localized in the nucleus; while in the GFP-transfected cells,GFP was localized in both the cytoplasm and nucleus. After OGD and H2O2 treatments, GFP/5-LOX was translocated to the nuclear membrane in 50.6% and 57.7% cells respectively. However,after NMDA treatment or in GFP-transfected cells, no translocation was observed. Wild-type 5-LOX was distributed in the nuclei and cytoplasm,and all the 3 treatments induced 5-LOX translocation to the nuclear membrane. Conclusion: In the PC12 cells stably transfected with GFP/5-LOX, GFP/5-LOX is primarily distributed in the nuclei; the OGD-,H2O2- and NMDA-induced 5-LOX translocation exhibits different properties.

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期刊信息
  • 《浙江大学学报:医学版》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国教育部
  • 主办单位:浙江大学
  • 主编:罗建红
  • 地址:杭州市天目山148号
  • 邮编:310007
  • 邮箱:zdxbyxb@zju.edu.cn
  • 电话:0571-88272797
  • 国际标准刊号:ISSN:1008-9292
  • 国内统一刊号:ISSN:33-1248/R
  • 邮发代号:32-2
  • 获奖情况:
  • 三次全国高校优秀自然科学学报评比先后荣获二等奖...
  • 国内外数据库收录:
  • 美国化学文摘(网络版),波兰哥白尼索引,荷兰文摘与引文数据库,美国生物医学检索系统,中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:6685