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目的:观察C6脑胶质瘤单次全脑放疗后早期血脑屏障(BBB)通透性的变化、超微结构特征及紧密连接蛋白Claudin-5的表达变化。方法:荷瘤鼠50只,随机分为假照射组和5、10、15、20Gy组各10只,行相应剂量全脑放射线单次照射,并于照射后24h处死,各5只分别行硝酸镧透射电镜示踪法观察肿瘤临近处(BAT)BBB通透性,免疫组化法测定Claudin-5的变化。结果:硝酸镧示踪电镜观察:假照射组、5Gy组荷瘤鼠BAT处见La3+渗出至局部基底膜,大部分分布在血管腔内;10Gy组BAT处可见La3+渗出至广泛基底膜及脑组织间隙,紧密连接开放;15、20Gy组BAT处均可见La3+渗出至基底膜、血管腔外、脑组织间隙内及细胞内。统计学分析硝酸镧渗出显示,20、15Gy>10Gy>5、0Gy,P<0.05。Clau-din-5表达变化:假照射组、5Gy组荷瘤鼠BAT处脑组织表达下降,仍呈中度表达;10Gy组BAT处可见较假照射组脑组织表达下降,呈轻中度表达,血管扭曲、变形与间断表达状态并存;15、20Gy组BAT处均可见较假照射组脑组织表达下降,呈轻度表达,沿血管间断表达,并见碎片状。统计学分析显示,20、15Gy>10Gy>5、0Gy,P<0.05。结论:荷瘤鼠全脑放疗后早期放射线损伤BBB的完整性发生在单次放疗剂量10～15Gy,且增加剂量BBB通透性无明显增加。
OBJECTIVE: To observe the changes of early BBB permeability, ultrastructural features and the expression of claudin-5 in C6 glioma after single whole-brain radiotherapy. METHODS: Fifty tumor-bearing mice were randomly divided into sham irradiation group and 10 rats in each of 5, 10, 15 and 20 Gy groups. The rats were irradiated with a single dose of whole brain radiation and killed 24 hours after irradiation. Five rats were treated with nitric acid The transmission electron microscope (TEM) was used to observe the permeability of BBB adjacent to the tumor (BAT). The change of Claudin-5 was detected by immunohistochemistry. Results: Lanthanum nitrate tracer electron microscopy: in sham-irradiated group, La3 + exuded to the local basement membrane in the 5Gy group of tumor-bearing mice, most of which were distributed in the vascular lumen. In the BAT of 10Gy group, La3 + exuded to the extensive basement membrane and Brain tissue gap, close connection open; 15,20 Gy group BAT Department can see La3 + exudation to the basement membrane, vascular extraluminal brain tissue space and intracellular. Statistical analysis of lanthanum nitrate exudation showed 20, 15 Gy> 10 Gy> 5,0 Gy, P <0.05. The expression of Clau-din-5 in the sham-irradiated group was lower than that in the 5Gy group (P <0.05). The expression of Clau-din-5 in the sham-irradiated group was lower than that in the 5Gy group Vascular distortions, deformation and intermittent expression state co-exist; 15, 20Gy group BAT can be seen compared with sham irradiation group decreased expression of brain tissue was mild expression along the vascular interrupted expression, and see the debris. Statistical analysis showed 20, 15 Gy> 10 Gy> 5,0 Gy, P <0.05. CONCLUSION: The integrity of BBB in early stage of radiation-induced TBI in tumor-bearing mice occurs at a single dose of 10-15 Gy, with no significant increase in BBB permeability at increasing doses.