Spautin-1 in improving anxiety-like behaviors in mice after traumatic brain injury through inhibiting astrocytic pyroptosis in the amygdala
10.3760/cma.j.cn115354-20211123-00759
- VernacularTitle:Spautin-1通过抑制杏仁核区星形胶质细胞焦亡改善小鼠TBI后焦虑样行为
- Author:
Huitao MIAO
1
;
Yonghan CHEN
;
Rongxin SONG
;
Zhiyou WU
;
Yue XIN
;
Jiexia WANG
;
Dongxue ZHANG
;
Limin ZHANG
Author Information
1. 河北医科大学附属沧州市中心医院神经外科，沧州 061000
- Keywords:
Traumatic brain injury;
Anxiety;
Autophagy;
Amygdala;
Pyroptosis
- From:
Chinese Journal of Neuromedicine
2022;21(6):553-562
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of Spautin-1 (an inhibitor of autophagy) on improving anxiety-like behaviors and its mechanism in mice after traumatic brain injury (TBI).Methods:Thirty-six C57BL/6 mice were randomly divided into sham-operated group, TBI group, and TBI+Spautin-1 group ( n=12); TBI models in the latter two groups were established by modified Feeney free fall epidural impingement method. Mice in TBI+Spautin-1 group were administered with Spautin-1 (2 μL, 10 mmol/L) into the lateral ventricle 10 min after modeling, but mice in the other two groups were only injected with same volume of solvent. Neuropathy Symptom Score (NSS) was used to evaluate the functions of motor, sensory and reflexes of mice on 1 st, 7 th and 14 th d of modeling. On 15 th and 16 th d of modeling, open field test and elevated plus maze test were used to evaluate the anxiety-like behaviors in mice. The number of Nissl bodies in the amygdala of mice was calculated by Nissl staining 16 d after modeling. The numbers of neuron specific nucleoprotein (NeuN) positive cells, interleukin (IL)-18 and IL-1β positive astrocytes in the amygdala were detected by immunofluorescent staining. Western blotting was used to detect the autophagy-and pyrotopic-associated protein expressions in the amygdala region of mice. Results:(1) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased NSS scores on 1 st and 7 th d of modeling ( P<0.05). (2) Open field test showed that as compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller number of crossing grids, significantly decreased percentage of time spending in the central zone ([central area residence time/total time] × 100%), significantly decreased percentage of frequencies entering into opening arm (OE) (OE/[OE+frequencies of entering closing arm]×100%) and opening arm time (OT) percentage (OT/[OT+time of closing arm]×100%); as compared with the TBI group, TBI+Spautin-1 group had significantly larger number of crossing grids, and significantly increased time percentage spending in the central zone, OE percentage, and OT percentage ( P<0.05). (3) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly smaller numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05); as compared with TBI group, TBI+Spautin-1 group had significantly larger numbers of Nissl bodies and NeuN positive cells in the amygdala of mice ( P<0.05). (4) As compared with the sham-operated group, the TBI group and TBI+Spautin-1 group had significantly increased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05); as compared with the TBI group, the TBI+Spautin-1 group had significantly decreased percentages of IL-1β and IL-18 positive astrocytes in amygdala of mice ( P<0.05). (5) As compared with sham-operated group, the TBI group and TBI+Spautin-1 group had significantly higher protein expressions of NOD-like receptor family protein 3 (NLRP3), activated cysteine aspartate protease-1 (Caspase-1), pore-forming protein D-N terminal fragment (GSDMD-N), ubiquitin specific peptidase (USP) 13 and B-lymphocytoma-2 interacting protein (Beclin1), and statistically higher ratio of microtubule-associated protein 1 light chain (LC)3 II/LC3 I ( P<0.05); as compared with TBI group, the TBI+Spautin-1 group had significantly decreased protein expressions of NLRP3, activated Caspase-1, GSDMD-N, USP13 and Beclin1 in the amygdala, and statistically lower ratio of LC3 II/LC3 I ( P<0.05). Conclusion:Spautin-1 improves the anxiety-like behaviors in mice after TBI, whose mechanism may be associated with the inhibition of astrocytic pyroptosis in the amygdala.
