Role of repressor element 1-silencing transcription factor/repressor element 1-silencing transcription factor coinhibitory factor in reparation of cortical neural axons after traumatic brain injury: an in vitro study
10.3760/cma.j.issn.1671-8925.2018.09.006
- VernacularTitle:REST/CoREST对TBI后皮层神经元轴突修复作用的体外研究
- Author:
Jun HE
1
;
Jie LIU
;
Jianke KUAI
;
Yuqin YE
;
Yongxiang YANG
;
Xiaosheng HE
Author Information
1. 710018,西安市第三医院麻醉科
- Keywords:
Traumatic brain injury;
Oxygen glucose deprivation;
Repressor element 1-silencing transcription factor;
Repressor element 1-silencing transcription factor coinhibitory factor
- From:
Chinese Journal of Neuromedicine
2018;17(9):897-904
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effects of repressor element 1-silencing transcription factor (REST)/REST coinhibitory factor (CoREST) on axonal regeneration and repairmen of mouse cortical neurons after traumatic brain injury (TBI).Methods (1) The primary cortical neurons were obtained from fetal C57BL/6 mice;one,three,5,7,9,and 11 d after cultivation,miR-124 expression was detected by quantitative-(q-) PCR.(2) Neurons cultured for 5 d were divided into miR-124 mimics group,blank control group,and miR-124 inhibitor group,and miR-124 mimics,nonsense control sequences and miR-124 inhibitor were transfected,respectively;0,6,12,24,48,and 72 h after transfection,miR-124 expression was detected by q-PCR.(3) Neurons cultured for 7 d were divided into blank control group Ⅰ,oxygen glucose deprivation (OGD) model group,up-regulated miR-124+OGD model group,and down-regulated miR-124+OGD model group,and neurons in the later two groups were transfected with miR-124 mimics and miR-124 inhibitor;48 h after transfection,OGD models in the later three groups were prepared;0,6,12,24,48,and 72 h after OGD,miR-124 expression was detected by q-PCR;GAP-43,REST and CoREST expressions were detected by Western blotting 48 h after OGD;the REST and CoREST expressions were measured by immunofluorescent staining 48 h after OGD.Results (1) One,three,5,and 7 d after cultivation,miR-124 expression gradually increased,and 7,9,and 11 d after cultivation,miR-124 expression gradually decreased,with significant differences (P＜0.05).(2) Twenty-four and 48 h after transfection,miR-124 expression in the miR-124 inhibitor group was significantly lower than that in the blank control group (P＜0.05);12,24,48 and 72 h after transfection,miR-124 expression in the miR-124 mimics group was significantly higher than that in the blank control group (P＜0.05),and peak level was noted at 48 h.(3) The miR-124 expression in the OGD model group was significantly higher than that in the blank control group Ⅰ at 12,24,48 and 72 h after OGD (P＜0.05),and peak level was noted at 48 h;0,6,12,24,48,and 72 h after OGD,the miR-124 expression in the up-regulated miR-124+OGD model group was significantly higher than that in the blank control group Ⅰ (P＜0.05),and peak level was noted at 48 h;Western blotting indicated that GAP-43 and CoREST gradually increased,and REST gradually decreased in blank control group Ⅰ,OGD model group and down-regulated miR-124+OGD model group,with significant differences (P＜0.05);neurons in the up-regulated miR-124+OGD model group had significantly lower GAP-43 and CoREST expressions,and significantly higher REST expression than those in the OGD model group (P＜0.05);the results of immunofluorescence staining were consistent with those of Western blotting.Conclusion REST/CoREST,as a pair regulator,may play a key role in the repairment and regeneration of neuron axons after TBI.
