Ceciliani Fabrizio, Soler Laura, Grilli Guido, Marques Andreia T, Giudice Chiara, Lecchi Cristina
Department of Veterinary Sciences and Public Health, Università di Milano, Via Celoria 10, 20133 Milano, Italy.
UMR PRC - URA - INRA Val de Loire, 37380 Nouzilly, France.
Vet Immunol Immunopathol. 2015 Nov 15;168(1-2):35-9. doi: 10.1016/j.vetimm.2015.08.004. Epub 2015 Aug 14.
In this article the localization of the acute phase protein Serum Amyloid A (SAA) in different depots of bovine adipose tissue (AT) and liver is reported. Quantitative (Real Time) PCR was paired to immunohistochemistry after the production of a specific polyclonal antibody. SAA's mRNA was found in all analyzed AT depots included in the present study, the AT located in the withers being the major source of SAA mRNA. A polyclonal antibody was raised against bovine SAA and was used to validate gene expression analyses. Western Blotting confirmed that SAA is present in all the seven adipose tissue depots include in the present experiment. Anti-SAA polyclonal antibody also stained diffusely adipocytes. In liver, intracytoplasmic immunolabeling was observed in hepatocytes. Staining was generally mild and not diffuse: negative hepatocytes were intermixed with positive ones. A positive intracytoplasmic immunostaining was occasionally observed in endothelial cells lining small blood vessels within AT septa and liver parenchyma. Our data confirm that bovine AT may provide an important source of SAA in healthy subjects. It remains to be determined which is the contribution of AT in the serum concentration of SAA.
本文报道了急性期蛋白血清淀粉样蛋白A（SAA）在牛脂肪组织（AT）和肝脏不同储存部位的定位。在制备特异性多克隆抗体后，将定量（实时）PCR与免疫组织化学相结合。在本研究纳入分析的所有AT储存部位均发现了SAA的mRNA，位于肩胛部的AT是SAA mRNA的主要来源。制备了针对牛SAA的多克隆抗体，并用于验证基因表达分析。蛋白质印迹法证实，在本实验纳入的所有七个脂肪组织储存部位均存在SAA。抗SAA多克隆抗体也能使脂肪细胞呈弥漫性染色。在肝脏中，在肝细胞内观察到胞浆内免疫标记。染色通常较淡且不呈弥漫性：阴性肝细胞与阳性肝细胞相互交织。在AT间隔和肝实质内的小血管内皮细胞中偶尔也观察到阳性胞浆免疫染色。我们的数据证实，在健康受试者中，牛AT可能是SAA的重要来源。AT对血清SAA浓度的贡献仍有待确定。
