Khoso Pervez Ahmed, Pan Tingru, Wan Na, Yang Zijiang, Liu Ci, Li Shu
College of Veterinary Medicine, Northeast Agricultural University, Harbin, 150030, People's Republic of China.
Biol Trace Elem Res. 2017 May;177(1):159-168. doi: 10.1007/s12011-016-0860-7. Epub 2016 Oct 15.
The aim of the present study was to investigate the effects of selenium (Se) deficiency on autophagy-related genes and on ultrastructural changes in the spleen, bursa of Fabricius, and thymus of chickens. The Se deficiency group was fed a basal diet containing Se at 0.033 mg/kg and the control group was fed the same basal diet containing Se at 0.15 mg/kg. The messenger RNA (mRNA) levels of the autophagy genes microtubule-associated protein 1 light chain 3 (LC3)-I, LC3-II, Beclin 1, dynein, autophagy associated gene 5 (ATG5), and target of rapamycin complex 1 (TORC1) were assessed using real-time qPCR. The protein levels of LC3-II, Beclin 1, and dynein were investigated using western blot analysis. Furthermore, the ultrastructure was observed using an electron microscope. The results indicated that spleen mRNA levels of LC3-I, LC3-II, Beclin 1, dynein, ATG5, and TORC1 and the protein levels of LC3-II, Beclin 1, and dynein were increased in the Se deficiency group compared with the control group. In the bursa of Fabricius, the mRNA levels of LC3-I, LC3-II, Beclin 1, dynein, ATG5, and TORC1 and the protein levels of Beclin 1 and dynein were increased; furthermore, the protein level of LC3-II was decreased in the Se deficiency group compared to the control group. In the thymus, the mRNA levels of LC3-I, Beclin 1, and ATG5 increased; the levels of LC3-II, dynein, and TORC1 were decreased; the protein level of Beclin 1 increased; and the levels of LC3-II and dynein decreased in the Se deficiency group compared to those in the control group. Further cellular morphological changes, such as autophagy vacuoles, autolysosomes, and lysosomal degradation, were observed in the spleen, bursa of Fabricius, and thymus of the Se-deficiency group. In summary, Se deficiency caused changes in autophagy-related genes, which increased the autophagic process and also caused structural damages to the immune organs of chickens.
本研究的目的是调查硒（Se）缺乏对鸡的自噬相关基因以及脾脏、法氏囊和胸腺超微结构变化的影响。缺硒组饲喂含硒量为0.033 mg/kg的基础日粮，对照组饲喂含硒量为0.15 mg/kg的相同基础日粮。使用实时定量PCR评估自噬基因微管相关蛋白1轻链3（LC3）-I、LC3-II、Beclin 1、动力蛋白、自噬相关基因5（ATG5）和雷帕霉素靶蛋白复合物1（TORC1）的信使核糖核酸（mRNA）水平。使用蛋白质印迹分析研究LC3-II、Beclin 1和动力蛋白的蛋白质水平。此外，使用电子显微镜观察超微结构。结果表明，与对照组相比，缺硒组脾脏中LC3-I、LC3-II、Beclin 1、动力蛋白、ATG5和TORC1的mRNA水平以及LC3-II、Beclin 1和动力蛋白的蛋白质水平均升高。在法氏囊中，LC3-I、LC3-II、Beclin 1、动力蛋白、ATG5和TORC1的mRNA水平以及Beclin 1和动力蛋白的蛋白质水平升高；此外，与对照组相比，缺硒组中LC3-II的蛋白质水平降低。在胸腺中，LC3-I、Beclin 1和ATG5的mRNA水平升高；LC3-II、动力蛋白和TORC1的水平降低；Beclin 1的蛋白质水平升高；与对照组相比，缺硒组中LC3-II和动力蛋白的水平降低。在缺硒组的脾脏、法氏囊和胸腺中观察到进一步的细胞形态变化，如自噬泡、自溶酶体和溶酶体降解。总之，硒缺乏导致自噬相关基因发生变化，这增加了自噬过程，也对鸡的免疫器官造成了结构损伤。
