Role of miR-205-3p in oncosis in astrocytes subjected to oxygen-glucose deprivation and restoration: relationship with AQP4
10.3760/cma.j.cn131073.20220329.00619
- VernacularTitle:miR-205-3p在氧糖剥夺-复氧复糖星形胶质细胞胀亡中的作用：与AQP4的关系
- Author:
Jingyan CHEN
1
;
Zhilin LIU
;
Hong WANG
;
Wenjie LIU
;
Kaiyue SHAN
;
Gaofeng ZHANG
;
Huailong CHEN
;
Mingshan WANG
;
Rui DONG
Author Information
1. 南京医科大学青岛临床医学院青岛市市立医院麻醉科，青岛 266071
- Keywords:
MicroRNAs;
Hypoxia;
Astrocytes;
Cell death;
Aquaporin 4
- From:
Chinese Journal of Anesthesiology
2022;42(6):734-738
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To evaluate the role of miR-205-3p in oncosis in astrocytes subjected to oxygen-glucose deprivation and restoration (OGD/R) and the relationship with aquaporin4 (AQP4).Methods:Primary astrocytes were cultured in vitro to the logarithmic growth phase and divided into 5 groups ( n=16 each) using a random number table method: control group (C group), OGD/R group (O group), OGD/R+ miR-205-3p mimic group (M group), OGD/R+ miR-205-3p inhibitor group (I group), and OGD/R+ negative control group (NC group). Cells were cultured routinely in C group.Cells were subjected to 4 h of oxygen-glucose deprivation in a 37℃ anaerobic incubator (containing 94% N 2, 1% O 2 and 5% CO 2) followed by restoration of O 2-glucose supply for 24 h in O group.Cells in M, I and NC groups were transfected with miR-205-3p mimic, miR-205-3p inhibitor and miR-205-3p negative control for 48 h, respectively, and then cells were subjected to 4 h of oxygen-glucose deprivation followed by restoration of O 2-glucose supply for 24 h. The cell viability was evaluated by CCK-8 assay, the cell injury and oncosis were analyzed by flow cytometry, the expression of AQP4 mRNA was detected by quantitative reverse transcription-polymerase chain reaction, and the expression of AQP4 and porimin was detected by Western blot. Results:Compared with C group, the expression of miR-205-3p was significantly down-regulated, the cell viability was decreased, the rates of cell injury and oncosis were increased, and the expression of AQP4 protein and mRNA and porimin was up-regulated in O group ( P<0.05). Compared with O group, the expression of miR-205-3p was significantly up-regulated, the cell viability was increased, the rates of cell injury and oncosis were decreased, and the expression of AQP4 protein and mRNA and porimin was down-regulated in M group, the expression of miR-205-3p was significantly down-regulated, the cell viability was decreased, the rates of cell injury and oncosis were increased, and the expression of AQP4 protein and mRNA and porimin was up-regulated in I group ( P<0.05), and no significant changes were found in NC group( P>0.05). Conclusions:miR-205-3p is involved in oncosis in astrocytes subjected to OGD/R, which is associated with regulation of AQP4 expression.
