Study of Time-Domain of Blood-brain Barrier Opened and Tight Junction Changed in Glioma Models after Whole-brain Irradiated with Single Dose
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摘要: 目的 观察C6脑胶质瘤15 Gy单次全脑放射线照射后血-脑屏障(blood-brain barrier,BBB)通透性的动态变化及紧密连接蛋白claudin-5 的表达变化，观察大鼠血-脑屏障(BBB)开放的时间阈。方法荷瘤鼠（建模后17天）60只，行15 Gy全脑放射线单次照射，并于照射后24 h、3 d、7 d、14 d、21 d、28 d各处死10只，各5只分别行硝酸镧（La+）透射电子显微镜示踪法观察肿瘤邻近处（brain adjacent to tumor,BAT）即距肿瘤边界2 mm以内BBB通透性及免疫组织化学法测定紧密连接蛋白claudin -5的变化研究。结果（1）照射后24 h、3 d硝酸镧渗出广泛，基底膜、血管腔外、脑组织间隙内及细胞内均可见；第7 d为La3+渗出减弱，基底膜及脑组织间隙内见细小的镧颗粒；第14 d为La3+渗出至局部基底膜，较前减少；第21 d、28 d均为La3+ 无渗出。统计学分析硝酸镧渗出程度显示：24 h和3 d>7 d>14 d>21 d和28 d（P<0.05）。（2）照射后24 h、3 d紧密连接蛋白claudin-5呈轻度表达，呈碎片状表达；照射后第7 d为轻中度表达，血管扭曲、变形；照射后第14 d为中度表达，血管扭曲、变形，但仍呈连续状态；照射后第21 d、28 d为中强度表达，出现连续与碎片表达并存。统计学分析claudin-5表达程度显示：24 h和3 d<7 d<14 d<21 d和28 d（P<0.05）。结论荷瘤鼠单次15 Gy照射后BBB开放，且2周后BBB通透性逐渐消失，表现出时间依赖性。Abstract: Objective To investigate the change in blood-brain barrier （BBB）permeability and ultrastructure of microvessle in C6 glioma and the tight junction protein expression of claudin-5，which were whole-brain irradiated with 15 Gy，and to explore the time-domain of BBB opened. Methods Sixty tumor-bearing rats (17th day after inoculation) were whole-brain irradiated with 15 Gy,and 10 rats were severally sacrificed at 24 h,3 d,7 d,14 d,21 d and 28 d after irradiation,5 rats were using transmission electron microscopy of exogenous lanthanum nitrate（La3+）-tracing to observe the BBB permeability of the tissues within 2 mm to the border of the tumor and using immunohistochemical method to detect the expression of claudin-5,respectively. Results (1)At 24 h,3 d after exposure,La3+ obviously exuded to the basement membrane（BM）,extra-vascular,the space inside the brain tissue and cells,a little remaining in the endovascular.At 7 d after exposure,La3+ obviously exuded to the basement membrane and brain tissue space less than that in the previous days.At 14d after exposure,La3+ exuded to the local basement membrane.At 21 d,28 d after exposure,La3+ exudation was disappeared.Statistical analysis showed that 24 h,3 d>7 d>14 d>21 d,28 d (P<0.05).(2)claudin-5 expression was mild at 24 h and 3 d after irradiation,showed debris-like expression.claudin-5 expression was light and moderate at 7 d after exposure,combined with deformation and continuous expression.It was moderate expression 14 d after exposure,showed vascular distortion and deformation,but was still continuous state.It was medium-intensity expression at 21 d and 28 d after irradiation,appeared to express both a row and debris.Statistical analysis showed that 24 h,3 d<7 d<14 d<21 d,28 d(P<0.05). Conclusion BBB is opened and gradually resuming 2 weeks after irradiated with 15Gy in C6 glioma which is in time-dependent manner.
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Key words:
- C6 glioma /
- Irradiation /
- Blood-brain barrier（BBB） /
- Ultrastucture /
- claudin-5
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