Influence of exosome-derived miR-124 on molecular expression related to axonal regeneration after mechanical damage to cortical neurons in mice
10.3760/cma.j.issn.1671-8925.2018.05.002
- VernacularTitle:外泌体源性miR-124对神经元损伤后轴突再生相关因子表达的影响
- Author:
Yongxiang YANG
1
;
Yuqin YE
;
Xinhong SU
;
Xin ZHANG
;
Chuiguang KONG
;
Wei BAI
;
Xiaosheng HE
Author Information
1. 空军军医大学西京医院神经外科
- Keywords:
miR-124;
Exosomes;
Cortical neuron injury;
Axonal regeneration
- From:
Chinese Journal of Neuromedicine
2018;17(5):440-444
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the influence of exosome-derived miR-124 on the molecular expression related to axonal regeneration after mechanical damage to cortical neurons in mice,aiming to provide experimental data for intervention in neurogenesis after traumatic brain injury (TBI).Methods The plasmid loaded with miR-124 was used to transfect the HEK293 cell line.The transfection effect was identified by real time Polymerase Chain Reaction (qPCR).The exosomes were isolated from the supematant of cultured transfected HEK293 cell line by the SBI isolation kit.The isolated exosomes were identified by electron microscopy and Western blotting,and the involved miR-124 in the exosomes was identified by qPCR.After the cortical neurons were isolated from the pregnant mice (14-17-day old) and cultured for 7 days,they were divided into 4 groups:control,damage,damage + exosomes without miR-124 and damage + exosomes with miR-124.The Petri dishes were manually scratched with a 10 μL plastic stylet needle to construct a mechanical damage in vitro in the latter 3 groups.The isolated exosomes without or with miR-124 were added into the cultured medium for culture for 72 h in the latter 2 groups,respectively.The expression ofmiR-124,NRP-1,Tau and Gap-43 was measured by qPCR and Western blotting respectively.Results The exosomes containing miR-124 were successfully obtained by plasmid transfection and the SBI isolation kit.The expression levels of miR-124,NRP-1 and Gap-43 in the damage + exosomes with miR-124 group were elevated significantly greater than in the other 3 groups (P＜0.05).The expression levels ofmiR-124,NRP-1 and Gap-43 in the damage group and damage + exosomes without miR-124 group were elevated significantly greater than in control group (P＜0.05).Conclusions The exosomes may transmit miR-124 to the cortical neurons in mice after mechanical damage and increase the expression ofmiR-124,NRP-1 and Gap-43 in the cortical neurons in mice.
