中国现代医学杂志2026，Vol.36Issue(11)：31-42,12.DOI:10.3969/j.issn.1005-8982.2026.11.005
EFhd2和ATG16L1在二乙酰吗啡致神经元自噬中的作用研究
Study on the role of EFhd2 and ATG16L1 in neuronal autophagy induced by diacetylmorphine
摘要
Abstract
Objective To explore the roles of EF-Hand domain family member 2(EFhd2)and Autophagy-related 16-like 1(ATG16L1)in diacetylmorphine-induced neuronal autophagy.Methods Twelve male SD rats were randomly divided into a control group and a model group,with six rats in each group.The model group was intraperitoneally injected with diacetylmorphine for 20 consecutive days to establish a diacetylmorphine addiction model,while the control group was injected with saline.Rat brain tissues were collected for proteomic sequencing to screen differential proteins;HE staining was used to observe morphological changes in brain tissue;and changes in autophagy protein expression levels were assessed.CCK-8 was used to determine the optimal concentration and changes in cell viability following diacetylmorphine intervention;Western blotting was used to detect changes in the levels of EFhd2,ATG16L1,and autophagy-related proteicontrolP62 and LC3;primary rat cortical neurocontrolwith EFhd2 knockdown were prepared and transfection efficiency was verified by Western blot;GFP-RFP-LC3 dual-fluorescence lentivirus was used to detect neuronal autophagic flux;the protein interaction sites were predicted and EFhd2-ATG16L1 interaction was validated by CO-IP.After activating ATG16L1 with Peretinoin,cell survival rate and autophagy levels were measured.Results Proteomics identified differential proteins,and among the significantly different proteins,EFhd2 was determined to be the primary research factor through interaction with the autophagy database.HE staining results showed that the brain tissue of the model group showed weaker staining compared with the control group,with a reduced number of nuclei.Western blot results indicated significant differences in the levels of P62,LC3Ⅱ/Ⅰ,EFhd2,and ATG16L1 between the controlgroup and model group(P<0.05);the control group had higher P62 levels than the model group(P<0.05),while LC3Ⅱ/Ⅰ,EFhd2,and ATG16L1 levels were lower than those in the model group.In vitro cell experiments demonstrated that after diacetylmorphine intervention on neuronal cells,variance analysis showed statistically significant differences in cell viability and autophagy-related protein expression,with cell viability markedly decreased compared with the NC group(P<0.05)and autophagy-related protein expression increased compared with the NC group(P<0.05).Following EFhd2 knockdown with the use of diacetylmorphine intervention(shEFhd2 DA group),variance analysis indicated significant differences in cell viability and autophagy protein levels,with cell viability significantly higher than the diacetylmorphine treatment group(DA group)(P<0.05)and autophagy protein expression lower compared with the DA group(P<0.05),indicating reduced autophagy levels relative to the DA group.Protein interaction prediction and CO-IP results showed an interaction between EFhd2 and ATG16L1.After activation of ATG16L1,variance analysis of cell viability showed statistically significant differences,with cell viability reduced and autophagy levels increased relative to the shEFhd2 DA group(P<0.05).Conclusion EFhd2 and ATG16L1 are involved in the neuronal autophagy process induced by diacetylmorphine.Modulating EFhd2 expression can reduce the level of diacetylmorphine-induced neuronal autophagy,providing a new target for the prevention and treatment of neurological diseases.关键词
二乙酰吗啡/自噬/EF-Hand结构域家族成员2/自噬相关蛋白16L1Key words
diacetylmorphine/autophagy/EF-Hand domain family member 2/autophagy-related protein 16L1分类
医药卫生引用本文复制引用
戴晨璐,刘静宇,苏丽萍,季敏,刘轩铭,程明,梁敏,蒲红伟..EFhd2和ATG16L1在二乙酰吗啡致神经元自噬中的作用研究[J].中国现代医学杂志,2026,36(11):31-42,12.基金项目
国家自然科学基金(82360256 （）
82460257) （）
广东省脑连接图谱重点实验室资助项目(2023B1212060055) （2023B1212060055）
