摘要
为研究鸡传染性法氏囊病毒（IBDV）感染对宿主免疫细胞线粒体代谢的影响和从代谢方面揭示致病机制,本试验以IBDV毒株感染DT40细胞,分别于病毒感染后24,48,72和96h取样,细胞计数并分离细胞线粒体,通过线粒体染色,检测线粒体蛋白含量,线粒体膜电位,线粒体ATP水平,肉碱脂酰转移酶和NADH-CoQ还原酶活性,线粒体脂酰-CoA和NADH、NAD^＋的含量及NADH/NAD^＋比值,初步评估IBDV感染对宿主细胞线粒体生物氧化的影响。结果表明,IBDV感染DT40细胞48h后,线粒体蛋白含量显著降低（P〈0.01）,肉碱脂酰转移酶和NADH-CoQ还原酶活性在感染后24h开始下降（P〈0.05）,48h后显著被抑制（P〈0.01）;脂酰-CoA和ATP含量在感染后48h显著降低（P〈0.01）,NADH/NAD＋比值显著增高（P〈0.01）。说明IBDV感染DT40细胞后不但对线粒体造成了损伤,而且脂酰CoA的转运和NADH呼吸链受到抑制,导致线粒体能量代谢障碍。本研究从代谢角度为进一步探索IBDV感染对鸡细胞的致病机理提供依据。
The purpose of this study was to determine the changes of mitochondrial energy metabolism in DT40 cells and its association with the pathogenicity of IBDV infection. DT40 cells were in- fected with IBDV isolate and the ceils were collected at 24,48,72 and 96 h post-inoculation. The mitochondrial fractions were extracted and mitochondrial protein levels and ATP contents were quantified by Bradford assay and phosphomolybdic acid hydrate eolorimetric method; acyl-CoA, NADH and NAD＋ were detected by colorimetric method. The activities of mitochondrial carnitine acyl transferase and NADH-CoQ reductases were detected by ELISA and colorimetric method. Results showed that the mitochondrial protein,acyl-CoA and ATP contents, the activities of carnitine acyl transferase and NADH-CoQ reductases in DT40 cells infected with IBDV were markedly decreased from 24 h （P〈0.05） and reached their minimum from 48 to 72 h post-inoculation （P〈 0.01） ;the ratio of NADH/NAD^＋ was markedly increased at 24 h compared with controls （P〈 0.05） ,peaking at 72 h post-inoculation （P〈0.01）. There was an association between NADH/ NAD^＋ ratio in mitochondria and an elevation in NADH-CoQ reductases activity. Taken together, our results demonstrated that mitochondrial energy metabolism disorder correlates with pathogenicity during IBDV infection. This study provides an incremental step in the understanding of the pathogenesis of IBDV.
作者
杨丹芳
李银聚
张春杰
余祖华
贾艳艳
何雷
王臣
韩海锋
YANG Dan-fang, LI Yin-ju , ZHANG Chun-jie, YU Zu-hua, JIA Yan-yan, HE Lei, WANG Chen, HAN Hai-feng(The Key Laboratory of Animal Disease and Public Security ,Henan University of Science and Technology, Luoyang, Henan 471003, Chin)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2018年第3期446-452,共7页
Chinese Journal of Veterinary Science
基金
河南省科技攻关资助项目(132102110117)
