Premium
[P2–185]: THE STUDY OF SUPPRESSION OF HYPOXIA‐INDUCED INFLAMMATION BY TIBETAN MEDICINE RATANASAMPIL IN MICROGLIA CELLS
Author(s) -
Zhong Xin,
Ni Junjun,
Meng Jie,
Nakanishi Hiroshi,
Wu Zhou,
Zhu Aiqin
Publication year - 2017
Publication title -
alzheimer's and dementia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.713
H-Index - 118
eISSN - 1552-5279
pISSN - 1552-5260
DOI - 10.1016/j.jalz.2017.06.836
Subject(s) - hypoxia (environmental) , microglia , viability assay , inflammation , pathogenesis , blot , mtt assay , cell , immunology , biology , pharmacology , chemistry , microbiology and biotechnology , medicine , oxygen , biochemistry , gene , organic chemistry
Background:Our previous studies have found that overexpression of 14-3-3z leads to SET retention in the cytoplasm. However, up-regulation of CKII increases SET Ser-9 phosphorylation and enhance SET retention in the cytoplasm. The relationship among 14-3-3z, SET and CKII remains unclear. Methods:AAV 14-3-3z was used to infect C57 mice. LSM and Co-immunoprecipitation was used to investigate the subcellular localization and interaction among SET, 14-3-3z, CKIIain brain slices. Contextual fear condition, Morris water maze test were used to detect the changes in behavior. LSM was used to verify SET subcellular localization in different area of brain slices. 64 array system was used to detect the LTP changes in living brain slices of hippocampal fromDG to CA1 region. Primary culture was employed to find the changes in neuronal dendrites and spines. Results:We found that 14-3-3zoverexpression induced SET localization in the cytoplasm while SET, 14-3-3z, CKIIa were interacted with each other. We observed that inhibition of CKII activity by TBB decreased phosphorylation level of SET at Ser-9 in 14-33ztransfectedHEK293/tau cell. si14-3-3zreduced SET pSer-9 in CKII transfectedHEK293/tau cell. Behavioral tests showed that si14-3-3z restored CKII-induced learning and memory impairments. CKIIa led to SET retention in the cytoplasm, while si14-3-3z promoted the SET nuclear importin. si14-3-3z treatment enhanced the reduction of LTP caused by CKIIa. Overexpression of CKIIa reduced neuronal dendrites length, dendritic branches and spine density, while si14-3-3z recovered these changes. We found that CKIIa phosphorylated SETand decreased the levels of synaptotagmin, synaptophysin, synapsin 1, p-synapsin 1, while si14-3-3zrestored these alterations. Conclusions: 14-3-3z interacts with both CKIIa and SET simultaneously, promotes CKII-catalyzed SET phosphorylation, induces neuronal dendritic impairment, LTP and learning and memory damage. si14-3-3z reduces CKIIa phosphorylating SET. Taken together, 14-3-3z recruiting CKIIa aggravates SET Ser-9 phosphorylation and induces downstream events of AD.