Apoptosis and leucocyte-endothelium interactions contribute to the delayed effects of cryotherapy on tumours in vivo
Apoptosis and leucocyte-endothelium interactions contribute to the delayed effects of cryotherapy on tumours in vivo
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DOI：
10.1007/s00403-002-0346-7
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发表时间：
2002-11-01
影响因子：
3
通讯作者：
Abels, C
中科院分区：
文献类型：
--
作者：
Schacht, V;Becker, K;Abels, C
在皮肤病学中，冷冻疗法通常用于治疗良性和恶性皮肤病变。然而，研究对微血管和随后的组织破坏的直接和延迟影响的时间过程缺乏。将无色素黑色素瘤（A-Mel-3）植入叙利亚金仓鼠（n=51）背侧皮褶腔。将肿瘤组织和正常组织冷冻至-20℃（冷却速度15℃/min）。采用活体荧光显微镜观察24小时内的微血管变化和白细胞-内皮相互作用。通过组织学和免疫组织化学观察坏死和凋亡的相对程度以及白细胞的积累。采用TUNEL法结合计算机辅助图像分析定量细胞凋亡。冷冻治疗后，毛细血管后小静脉和肿瘤血管的红细胞速度（RBCV）降低，而肿瘤血管的功能血管密度（FVD）与毛细血管后小静脉相比显著降低。白细胞-内皮相互作用首先在正常组织和肿瘤组织中增加，24h后白细胞在靠近肿瘤边缘的正常组织中积累。冻疮在冷冻后直接引起坏死，并在整个观察期内保持不变。相比之下，在冷冻治疗后24小时，肿瘤组织周围的细胞凋亡增加。综上所述，冷冻疗法对组织的破坏不仅是由直接坏死和微血管淤滞引起的，还包括炎症浸润和随后的细胞凋亡。这可能是关于免疫反应产生的一个重要发现。
In dermatology, cryotherapy is commonly used to treat benign and malignant skin lesions. However, studies investigating the time-course of the direct and delayed effects on the microvasculature and subsequent tissue destruction are lacking. Amelanotic melanomas (A-Mel-3) were implanted into the dorsal skinfold chamber of Syrian Golden hamsters (n=51). Tumour and normal tissue were frozen to -20degreesC (cooling rate 15degreesC/min). Intravital fluorescence microscopy was performed to assess microvascular changes and leucocyte-endothelium interactions up to 24 h. The relative degrees of necrosis and apoptosis and the accumulation of leucocytes were investigated by histology and immnuohistochemistry. Apoptosis was quantified using the TUNEL assay in combination with computer-assisted image analysis. After cryotherapy, red blood cell velocity (RBCV) decreased in postcapillary venules and tumour vessels, whereas functional vessel density (FVD) was significantly reduced in tumour vessels as compared with postcapillary venules. Leucocyte-endothelium interaction increased first in normal and tumour tissue, and then after 24 h leucocytes accumulated in normal tissue close to the tumour margin. Necrosis was induced in the cryolesions directly after freezing and remained constant over the entire observation period. In contrast, apoptosis increased in the periphery of the tumour tissue up to 24 h following cryotherapy. In conclusion, tissue destruction by cryotherapy is not only induced by direct necrosis and microvascular stasis, but also by the inflammatory infiltrate and subsequent apoptosis. This could be an important finding regarding the generation of an immune response.