Croce Anna Cleta, De Simone Uliana, Vairetti Mariapia, Ferrigno Andrea, Boncompagni Eleonora, Freitas Isabel, Bottiroli Giovanni
IGM-CNR Histochemistry and Cytometry Section and Department of Animal Biology, University of Pavia, Piazza Botta, 10, 27100, Pavia, Italy.
Photochem Photobiol Sci. 2008 Sep;7(9):1046-53. doi: 10.1039/b804836c. Epub 2008 Jul 24.
Autofluorescence spectroscopy is a promising and powerful approach for an in vivo, real time characterization of liver functional properties. In this work, preliminary results on the dependence of liver autofluorescence parameters on the nutritional status are reported, with particular attention to vitamin A and lipid accumulation in liver tissue. Normally fed and 24 h starving rats were used as animal models. Histochemical and autofluorescence analysis showed that lipids and vitamin A colocalize in the liver parenchyma. Fasting condition results in a parallel increase in both lipids and vitamin A. Autofluorescence imaging and microspectrofluorometric analysis carried out on unfixed, unstained tissue sections under 366 nm excitation, evidenced differences in both spectral shape and response to continuous irradiation between liver biopsies from fed and starving rats. As to photobleaching, in particular, fitting analysis evidenced a reduction of about 85% of the signal attributable solely to vitamin A during the first 10 s of irradiation. The tissue whole emission measured in fed and starving rat livers exhibited reductions of about 35% and 52%, respectively, that are closely related to vitamin A contents. The findings open interesting perspectives for the set up of an in situ, real time diagnostic procedure for the assessment of liver lipid accumulation, exploiting the photophysical properties of vitamin A.
自体荧光光谱法是一种用于体内实时表征肝脏功能特性的很有前景且强大的方法。在这项工作中,报告了肝脏自体荧光参数对营养状况依赖性的初步结果,特别关注肝脏组织中维生素A和脂质的积累。正常喂食和饥饿24小时的大鼠被用作动物模型。组织化学和自体荧光分析表明,脂质和维生素A在肝实质中共定位。禁食状态导致脂质和维生素A同时增加。在366nm激发下对未固定、未染色的组织切片进行的自体荧光成像和显微分光荧光分析表明,喂食和饥饿大鼠肝脏活检组织在光谱形状和对连续照射的响应方面均存在差异。特别是关于光漂白,拟合分析表明,在照射的前10秒内,仅归因于维生素A的信号减少了约85%。喂食和饥饿大鼠肝脏中测得的组织整体发射分别减少了约35%和52%,这与维生素A含量密切相关。这些发现为利用维生素A的光物理特性建立一种用于评估肝脏脂质积累的原位实时诊断程序开辟了有趣的前景。