摘要
目的评价在体生物发光成像术(bioluminescence imaging,BLI)监测小鼠体内骨髓来源细胞(bone marrow-derived cells,BMCs)参与脉络膜新生血管(choroidal neovascularization,CNV)形成过程的可行性,为BLI研究CNV发生机制奠定基础。方法将荧光素酶-绿色荧光蛋白(Fluc-GFP)双转基因小鼠的骨髓移植到野生型C57小鼠,将嵌合度>85%的嵌合体小鼠分为两组:实验组利用532nm激光建立小鼠CNV模型,对照组不行CNV建模。于CNV建模后1d、3d、7d、14d、21d和28d,利用小动物成像系统对两组小鼠进行BLI观察,高灵敏度CCD采集图像,活体成像软件系统对选取的眼部光学统计区域进行分析。结果建模后第1天,实验组小鼠即有大量BMCs趋化至CNV部位,前3d趋化至CNV部位的BMCs数量增长最快,第7天病变部位BMCs数量达峰值,第14天病变部位BMCs数量降至最低,随后稍有增加,第28天以后保持稳定。对照组小鼠在相应时间点眼内BMCs数量较为稳定,明显低于实验组小鼠(P<0.05)。结论利用BLI可以实时动态示踪体内BMCs向CNV部位的趋化及其在CNV内转归,无创、便捷,为研究CNV发生机制提供了一种新的客观的辅助方法。
Objeetive To evaluate the feasibility of using in vivo bioluminescence imaging （BLI） to observe the dynamic conduct of bone marrow-derived cells （BMCs） in choroidal neovascularization（CNV）, and pave the way for using BLI to investigate the mechanism of CNV. Methods Chimeric mice were developed by transplanting bone marrow from genetically modified mice having Fluc and green fluorescent protein（GFP） reporter genes to wild-type adult C57 mice. All chimeric mice that had a chimerism degree higher than 85% were divided into control group and experimental group. CNV model was established in the experimental group, the control group was unchanged. At the 1st day,3rd day,7th day,14th day,21st day and 28th day after model- ing, BLI observation was performed on the two groups using IVIS Kinetics system,image capture was preformed with high sensitive CCD. Then regions of interest were analyzed using Living Imaging Version 4.0 software, Results At the 1st day after modeling, a large number of BMCs were found at the CNV area; The number of BMCs found at the CNV area increased fastest during the flrst 3 days and it peaked at the 7th day;At the 14th day,it bottomed, and increased a little afterwards; It remained stable at the 28th day. The numbers of BMCs found at the CNV area of the control group remained stable at each points-in-time and were significantly smaller that of the experimental group（P 〈 0.05 ）. Conelusion In vivo BLI could trace the transition of BMCs at the CNV area in a dynamic and real-time manner,noninvasively and conveniently, which is a new method of investigating the mechanism of CNV.
出处
《眼科新进展》
CAS
北大核心
2013年第5期401-404,409,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:81070748
81200708)
国家重点基础研究发展计划(编号:2011CB510200)
德国洪堡基金会仪器设备捐赠基金资助~~
关键词
脉络膜新生血管
骨髓来源细胞
在体生物发光成像术
choroidal neovascularization
bone marrow-derived cells
in vivo bi-oluminescence imaging
