Low-Frequency Ultrasound Irradiation Increases Blood-Tumor Barrier Permeability by Transcellular Pathway in a Rat Glioma Model
Low-Frequency Ultrasound Irradiation Increases Blood-Tumor Barrier Permeability by Transcellular Pathway in a Rat Glioma Model
复制标题
低频超声照射通过大鼠胶质瘤模型中的跨细胞途径增加血液肿瘤屏障的渗透性
DOI：
10.1007/s12031-012-9770-0
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发表时间：
2012-09-01
影响因子：
3.1
通讯作者：
Xue, Yi-xue
中科院分区：
文献类型：
--
作者：
Xia, Chun-yi;Liu, Yun-hui;Xue, Yi-xue
低频超声(LFU)在一定声强下可无创、可逆地增加血脑屏障通透性。本研究旨在探讨LFU照射对大鼠C6脑胶质瘤模型血肿瘤屏障通透性的影响及其可能机制。本研究采用伊文思蓝染色和H&E染色，探讨了LFU在不损伤正常脑组织的情况下打开BTB的最佳参数。用透射电子显微镜观察脑瘤和脑胶质瘤微血管内皮细胞吞噬小泡数量的变化。免疫印迹法检测酪氨酸激酶Src、小窝蛋白-1和小窝蛋白-2的磷酸化。免疫组织化学和免疫荧光染色、RT-PCR和Western印迹检测小窝蛋白的分布和表达水平。我们的研究结果表明，在大鼠C6脑胶质瘤模型中，LFU频率为1 MHz，功率为12 mW，照射时间为20 S时，血脑屏障通透性呈暂时性升高，1.5h达高峰，12h后下降至正常水平。在胶质瘤模型大鼠脑胶质瘤微血管内皮细胞中，LFU照射后吞噬泡密度明显增加。免疫组织化学和免疫荧光染色显示小窝蛋白-1和小窝蛋白-2共定位于胶质瘤微血管内皮细胞和胶质瘤细胞中。LFU照射后，小窝蛋白-1和小窝蛋白-2的mRNA和蛋白表达水平上调，1.5h达到峰值，12h恢复正常。这些结果表明，LFU照射通过促进胶质瘤微血管内皮细胞的跨细胞转运而增加BTB的通透性。酪氨酸激酶Src、小窝蛋白-1、小窝蛋白-2的磷酸化以及小窝蛋白-1、小窝蛋白-2的上调参与了LFU诱导的小窝介导的内吞作用。
Low-frequency ultrasound (LFU) irradiation under certain acoustic intensity can increase blood-brain barrier permeability non-invasively and reversibly. The aim of this study was to find out the effect of LFU irradiation on blood-tumor barrier (BTB) permeability in rat C6 glioma model and the possible mechanism. In this research, Evans blue and H&E staining were used to evaluate the optimal parameter of LFU to open the BTB without damaging the normal brain tissue. Transmission electron microscopy was used to observe the changes of the number of pinocytotic vesicles in cerebral or glioma microvascular endothelial cells. The phosphorylation of tyrosine kinase Src, caveolin-1, and caveolin-2 was detected by western blot. The distribution and expressing levels of caveolae proteins, caveolin-1 and caveolin-2, were detected by immunohistochemical and immunofluorescent staining, RT-PCR, and western blot. Our research data showed that, in rat C6 glioma model, LFU irradiation at a frequency of 1 MHz, a power of 12 mW, and exposure time of 20 s induced the increase of BTB permeability temporally, which reached a peak at 1.5 h, then decreased and restored to normal level at 12 h after LFU irradiation. In the glioma microvascular endothelial cells of rat glioma model, LFU irradiation induced a significant increase of the pinocytotic vesicles' density. The phosphorylation of Src, caveolin-1, and caveolin-2 began to increase at 0.5 h and reached a maximum at 1 h. Immunohistochemical and immunofluorescent staining showed that caveolin-1 and caveolin-2 were co-localized in the glioma microvascular endothelial cells and glioma cells. The mRNA and protein expression levels of caveolin-1 and caveolin-2 were up-regulated, reached the peak value at 1.5 h, and re-normalized at 12 h after LFU irradiation. These results demonstrated that LFU irradiation increased BTB permeability by promoting transcellular transport in glioma microvascular endothelial cells. The phosphorylation of tyrosine kinase Src, caveolin-1, caveolin-2 and up-regulation of caveolin-1 and caveolin-2 were involved in LFU-induced caveolae-mediated endocytosis.