摘要
为研究VD_(3)影响亚临床低钙血症奶牛中性粒细胞内Ca^(2+)浓度对LC3自噬作用的影响,采集奶牛外周血分离提取中性粒细胞;Western blot技术对细胞LC3自噬途径相关蛋白(LC3-Ⅱ、P62、ATG5)丰度检测;通过流式细胞术检测中性粒细胞胞内Ca^(2+)浓度水平;利用激光共聚焦技术检测细胞LC3自噬体。通过添加VD_(3)作用影响细胞内Ca^(2+)浓度水平,探究VD_(3)促进细胞钙吸收影响LC3自噬功能。Western blot检测显示亚临床低钙血症奶牛中性粒细胞LC3、ATG5蛋白丰度下降,P62蛋白丰度上升,结果表明,亚低钙血症奶牛LC3自噬功能减弱。VD_(3)处理后,流式细胞术结果显示VD_(3)促进中性粒细胞内Ca^(2+)浓度水平上升。Western blot结果显示,健康奶牛和亚临床低钙血症奶牛中性粒细胞在VD_(3)作用下LC3自噬功能增强,说明VD_(3)促进中性粒细胞内Ca^(2+)浓度水平继而影响LC3自噬功能。
To investigate the effect of VD3 on the concentration of Ca^(2+)in neutrophils of subclinical hypocalcemia dairy cows and the autophagy of LC3,peripheral blood of dairy cows was collected and neutrophils were isolated and extracted.Western blot was used to detect the abundance of LC3 autophagy pathway-related proteins(LC3-Ⅱ,P62,ATG5)in cells.The intracellular Ca^(2+)concen-tration of neutrophils was detected by flow cytometry.The LC3 autophagosome was detected by la-ser confocal technology.By adding VD3 to affect intracellular Ca^(2+)concentration levels,the effects of vitamin D3 on calcium absorption and LC3 autophagy function were investigated.Western blot result showed that the protein abundances of LC3 and ATG5 in neutrophils of subclinical hypocal-cemia dairy cows decreased,while the protein abundance of p62 increased,indicating that the auto-phagy function of LC3 in subclinical hypocalcemia dairy cows was weakened.After VD3 treatment,the flow cytometry results showed that VD3 promoted the increase of Ca^(2+)concentration levels in neutrophils.The Western blot results showed that the LC3 autophagy function of neutrophils from healthy cows and subclinical hypocalcemia cows was enhanced under the action of VD3,indicating that VD3 promoted the concentration of Ca^(2+)in neutrophils and then affected the LC3 autophagy function.
作者
吕鑫泉
李思瑶
张冰冰
徐闯
杨威
LYU Xinquan;LI Siyao;ZHANG Bingbing;XU Chuang;YANG Wei(Collegeof LifeScience and Technology,Heilongjiang Bayi Agricultural University,Daqing,Heilongjiang 163000 China;College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611100,China;College of Animal Science and Technology,China Agricultural University,Beijing 100193,China)
出处
《中国兽医学报》
北大核心
2025年第6期1268-1272,共5页
Chinese Journal of Veterinary Science
基金
国家重点研发计划资助项目(2023YFD1801100)。
