[1]骆 杨 徐义昌 徐善水.Ad-GFP转染兔离体基底动脉环的研究[J].中国临床神经外科杂志,2015,(10):621-623.[doi:10.13798/j.issn.1009-153X.2015.10.014]
 LUO Yang,XU Yi-chang,XU Shan-shui..Preliminary research of transfection of Ad-GFP into rabbit basal artery ring in vitro[J].,2015,(10):621-623.[doi:10.13798/j.issn.1009-153X.2015.10.014]
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Ad-GFP转染兔离体基底动脉环的研究()
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《中国临床神经外科杂志》[ISSN:1009-153X/CN:42-1603/TN]

卷:
期数:
2015年10期
页码:
621-623
栏目:
论著
出版日期:
2015-10-30

文章信息/Info

Title:
Preliminary research of transfection of Ad-GFP into rabbit basal artery ring in vitro
文章编号:
1009-153X(2015)10-0621-03
作者:
骆 杨 徐义昌 徐善水
242000 安徽,宣城市人民医院神经外科(骆 杨、徐义昌);241001 安徽芜湖,皖南医学院弋矶山医院神经外科(徐善水)
Author(s):
LUO Yang1 XU Yi-chang1 XU Shan-shui2.
1. Department of Neurosurgery, People's Hospital of Xuan Cheng City, Xuancheng Anhui 242000, China;
2. Department of Neurosurgery, Yijishan Hospital, Wannan Medical College, Wuhu 241001, China
关键词:
基底动脉环体外培养基因转染绿色荧光蛋白新西兰纯种大白兔
Keywords:
Culture in vitro Basilar artery rings Endothelial cell Green fluorescent protein
分类号:
Q 789; Q 786
DOI:
10.13798/j.issn.1009-153X.2015.10.014
文献标志码:
A
摘要:
目的 通过将将携带有绿色荧光蛋白基因的重组腺病毒(Ad-GFP)转染兔离体基底动脉环,观察GFP表达部位及表达量的变化。方法 获取兔基底动脉环,置入含DMEM的96孔板中;加入1.8×106 pfu/μl的Ad-GFP转染1 h;转染后即刻3、5、7 d行冰冻切片,荧光显微镜观察亮绿色物质。另外,将不同病毒浓度(分别为1.2×105、1.2×106 、1.8×106和2.4×106 pfu/μl)转染离体血管环1 h后,培养3 d后行冰冻切片,用荧光显微镜观察结果。结果 荧光显微镜下观察发现,基底动脉环管壁各层均表达GFP,但主要位于血管内皮层,1 000倍下在内皮细胞内中明确观察到荧光颗粒。1.8×106 pfu/μl的Ad-GFP转染后发现,GFP在转染3 d后表达最明显,5 d次之,而7 d后表达最弱;不同浓度Ad-GFP转染3 d后发现,浓度为1.2×105 pfu/μl时,管壁有较弱绿色荧光,浓度不断升高亮度有所增强,浓度为1.8×106 pfu/μl时达到最高峰,浓度继续增加时荧光亮度未见进一步增强。结论 Ad-GFP可以成功转染入离体血管环中,以内皮层为主;3 d为病毒转染后最佳培养时间。
Abstract:
Objective To observe the expression sites and the quantity of the green fluorescent protein (GFP) in order to provide the base for recombinant adenovirus endothelial nitrogen oxide synthase transfection in vitro and in vivo. Methods The basilar artery rings derived from the rabbits were cultured in the media containing the same concentration of recombinant adenovirus encoding green fluorescent protein (Ad-GFP) for 0, 3, 5 and 7 days respectively and in the media containing the different concentrations of Ad-GFP for 3 days respectively. The expressions of GFP in the basilar artery rings were observed in all groups. Results The GFP could be detected in all the vascular walls, and the most significant expression was in the endothelia. When the concentration of Ad-GFP in the media increased, GFP expression quality also increased. The unchanged expression was not observed until the concentration of Ad-GFP in the media arrived at 1.8×106 pfu/μl. The expression of GFP were the strongest and the weakest respectively in 3 and 7 days groups among all the groups in which the basilar artery rings were cultured in the media containing the same concentration of Ad-GFP for different days. Conclusions Ad-GFP may be successfully transfected into the vascular walls, especially in the endothelia. It is suggested that the expression of GFP is best in the basilar artery rings which are cultured for 3 days in the medium with Ad-GFP concentration of 1.8×106 pfu/μl.

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更新日期/Last Update: 2015-10-30