Альманах клинической медицины. 2018; 46: 682-689
Роль ультраструктурных изменений гематоэнцефалического барьера в развитии радиорезистентности глиобластомы головного мозга
https://doi.org/10.18786/2072-0505-2018-46-7-682-689Аннотация
Актуальность. Глиобластома (ГБ) – наиболее часто диагностируемая опухоль головного мозга, при лечении которой используются адъювантные методы лечения, включая лучевую терапию. Причиной высокой вероятности возникновения местного рецидива ГБ считается ее радиорезистентность, обусловленная в том числе развитием гипоксии в результате нарушения проницаемости гематоэнцефалического барьера в сосудах ГБ и перитуморальной зоны (ПЗ).
Цель – изучить патогенез гипоксии в области резидуальной ГБ на основании данных об изменениях морфологических элементов сосудистой стенки капилляров, формирующих гематоэнцефалический барьер в капиллярах ГБ и ПЗ.
Материал и методы. Морфологический материал был получен в процессе удаления ГБ головного мозга у 5 пациентов. Подготовку материала для исследования методом трансмиссионной электронной микроскопии осуществляли по стандартной методике с фиксацией образцов в 2% растворе глутаральдегида на фосфатном буфере, постфиксацией тетраоксидом осмия, заливкой в смесь эпона и аралдита и контрастированием ультратонких срезов уранилацетатом и цитратом свинца. В двух группах капилляров – ГБ (n = 38) и ПЗ (n = 32) – оценивали наличие изменений в клеточных элементах капилляров (вакуолизация митохондрии и вакуолизация эндоплазматического ретикулума в эндотелиоците, периците и астроците), а также состояние неклеточного элемента капиллярной стенки – базальной мембраны.
Результаты. Выявлено наличие характерных для апоптоза и онкоза изменений в таких клеточных элементах, формирующих гематоэнцефалический барьер в капиллярах ГБ и ПЗ, как эндотелиоциты и перициты. Однако в капиллярах ГБ такие изменения выявлялись значительно чаще (р < 0,001). Только в половине (52,6%) капилляров ГБ обнаружен перикапиллярный астроцитарный слой, находящийся в состоянии отека. В остальных случаях астроцитарные отростки либо визуализировались в виде отдельных структурных элементов (13,2%), либо вообще не были обнаружены (34,2%). В капиллярах ПЗ астроцитарный слой имел место во всех наблюдениях, его отек отмечен в 68,8% капилляров, при этом тотальный – только в 25%. Расширение базальной мембраны отмечено в подавляющем большинстве (89,5%) капилляров ГБ и только в 25% капилляров ПЗ (р < 0,001).
Заключение. Обнаруженные изменения клеточных элементов капилляров ГБ, обусловливающие формирование перитуморального отека и, как следствие, возникновение гипоксии, с высокой степенью вероятности служат причиной радиорезистентности резидуальной ГБ.
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Almanac of Clinical Medicine. 2018; 46: 682-689
The role of ultrastructural abnormalities of the blood-brain barrier in the development of brain glioblastoma radioresistance
https://doi.org/10.18786/2072-0505-2018-46-7-682-689Abstract
Background: Glioblastoma (GB) is the most commonly diagnosed brain tumor. Its management involves adjuvant therapies, such as radiation. The cause of high probability of GB local relapse is its radioresistance related to hypoxia arising from abnormal blood-brain barrier permeability in GB vessels and in the peritumoral zone (PZ).
Aim: To study pathophysiology of hypoxia in the residual GB based on the abnormalities of the morphological elements of the capillary walls building up the blood-brain barrier in GB and PZ capillaries.
Materials and methods: Samples for morphological evaluation were taken during surgery for GB in 5 patients. The samples were prepared for transmission electron microscopy according to the standard technique with fixation in 2% glutaraldehyde in phosphate buffer, post-fixation with osmium tetroxide, embedding in the epon-araldite mixture, and contrast staining of ultrathin sections with uranylacetate and lead citrate. Abnormalities of the capillary cells (mitochondrial vacuolization and vacuolization of endoplasmic reticulum in endothelial cells, pericytes and astrocytes), as well as of the acellular element of the capillary wall, i.e. basement membrane, were assessed in two groups of capillaries – those of GB (n = 38) and those of PZ (n = 32).
Results: Abnormalities characteristic for apoptosis and oncosis were found in the cells of the GB and PZ capillaries of the blood-brain barrier, such as endothelial cells and pericytes. However, in the GB capillaries these abnormalities were signifcantly more frequent (р < 0.001). Only half (52.6%) of the GB capillaries had an edematous pericapillary astrocyte layer. In all other capillaries, astrocyte sprouts either were visualized as separate morphological elements (13.2%) or were not visualized at all (34.2%). All PZ capillaries had the astrocyte layer, being edematous in 68.8% of the capillaries and totally edematous only in 25%. Thickened basement membrane was found in the vast majority (89.5%) of the GB capillaries and only in 25% of the PZ capillaries (р < 0.001).
Conclusion: Findings of abnormal cell elements in the GB capillaries leading to peritumoral edema and consequent hypoxia are highly likely to be the cause of the remnant GB radioresistance.References
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24. Wesseling P, Schlingemann RO, Rietveld FJ, Link M, Burger PC, Ruiter DJ. Early and extensive contribution of pericytes/vascular smooth muscle cells to microvascular proliferation in glioblastoma multiforme: an immuno-light and immuno-electron microscopic study. J Neuropathol Exp Neurol. 1995;54(3):304–10.
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27. Castejón OJ. Ultrastructural alterations of human cortical capillary basement membrane in human brain oedema. Folia Neuropathol. 2014;52(1):10–21. doi: 10.5114/fn.2014.41740.
28. Castejón OJ, Castejón HV, Zavala M, Sánchez ME, Díaz M. A light and electron microscopic study of oedematous human cerebral cortex in two patients with post-traumatic seizures. Brain Inj. 2002;16(4):331–46. doi: 10.1080/02699050110088209.
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