Effect and mechanism of lncRNA HCP5 on neurocyte apoptosis induced by oxygen-glucose deprivation/reoxygenation
10.3760/cma.j.cn371468-20230214-00062
- VernacularTitle:lncRNA HCP5对氧糖剥夺/复氧诱导的神经细胞凋亡的影响及其机制
- Author:
Shanshan WANG
1
;
Junjie LI
;
Zhanping GUO
;
Ning ZHANG
Author Information
1. 新乡市中心医院（新乡医学院第四临床学院）检验科，新乡 453000
- Keywords:
lncRNA HCP5;
miR-525-5p;
B-cell translocation gene 2;
Oxygen-glucose deprivation/reoxygenation;
Nerve cells;
Apoptosis;
Oxidative stress
- From:
Chinese Journal of Behavioral Medicine and Brain Science
2023;32(9):794-801
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of long non-coding RNA (lncRNA) human histocompatibility leukocyte antigen complex P5 (HCP5) on the neuronal injury induced by oxygen glucose deprivation/reoxygenation (OGD/R), and to analyze its potential mechanism.Methods:SH-SY5Y cells were divided into control group (CON group, normal medium culture), model group (Model group, OGD/R), interference control group (si-NC group, OGD/R after HCP5 small interfering RNA negative control (si-NC)), HCP5 interference group (si-HCP5 group, OGD/R after HCP5 small interfering RNA (si-HCP5)), HCP5 interference+ inhibitor control group (si-HCP5+ anti-NC group, OGD/R after transfection of si-HCP5, miR-525-5p inhibitor negative control (anti-NC)), HCP5 interference+ miR-525-5p inhibitor group (si-HCP5+ anti-miR-525-5p group, OGD/R after transfection of si-HCP5, miR-525-5p inhibitor). qRT-PCR was used to detect the expression of lncRNA HCP5 and miR-525-5p in cells.The activity of SH-SY5Y cells was detected by MTT.The level of reactive oxygen species (ROS) in the cells was detected by fluorescent probe of dichlorofluorescein diacetate (DCFH-DA). The apoptosis of SH-SY5Y cells was detected by flow cytometry.Western blot was used to detect the expression of BTG2, Bcl-2 related X protein (Bax), B lymphocyte tumor 2 (Bcl-2) and cleaved caspase-3 protein.SPSS 25.0 software was used to analyze the data, and one-way ANOVA was used for comparison between multiple groups, and SNK- q test was used for further comparison between two groups. Results:There were statistically significant differences in lncRNA HCP5, miR-525-5p RNA levels and BTG2 protein expression levels among the 6 groups ( F=28.853, 59.241, 13.731, all P<0.001). Compared with the CON group, the Model group had higher level of lncRNA HCP5, lower level of miR-525-5p, and higher level of BTG2 protein (all P<0.05). Compared with the Model group, the si-HCP5 group had lower level of lncRNA HCP5, higher level of miR-525-5p, and lower level of BTG2 protein (all P<0.05). Compared with the si-HCP5+ anti-NC group, the si-HCP5+ anti-miR-525-5p group had higher level of lncRNA HCP5, lower level of miR-525-5p, and higher level of BTG2 protein (all P<0.05). There were statistically significant differences in cell activity and ROS levels among the six groups of cells ( F=16.180, 59.950, both P<0.001). The cell activity of the Model group was lower than that of the CON group (0.33±0.12, 0.63±0.11) ( P<0.05), and the ROS level was higher than that of the CON group (224.62±23.27, 100.00±0.00) ( P<0.05). The cell activity of the si-HCP5+ anti-miR-525-5p group was lower than that of the si-HCP5+ anti-NC group (0.38±0.08, 0.58±0.08) ( P<0.05), and the ROS level was higher than that of the si-HCP5+ anti-NC group (207.83±19.39, 135.27±14.36) ( P<0.05). There were statistically significant differences in the apoptosis rate and expression levels of apoptotic proteins Bcl-2, Bax, and cleared Caspase-3 among the six groups of cells ( F=27.994, 29.660, 45.000, 52.983, all P<0.001). There were no statistically significant difference in Bax, Bcl-2, cleared Caspase-3 protein levels, and apoptosis rate in SH-SY5Y cells between the Model group and the si-NC group, as well as between the si-HCP5 group and the si-HCP5+ anti-NC group (all P>0.05). Compared with the CON group, the apoptosis rate, levels of Bax and cleared Casase-3 protein in the Model group were significantly upregulated (all P<0.05), while the Bcl-2 protein level was significantly downregulated ( P<0.05). Compared with the Model group and si-NC group, the si-HCP5 group showed significant downregulation of cell apoptosis rate and levels of Bax and cleared Caspase-3 protein (all P<0.05), while the Bcl-2 protein level was upregulated ( P<0.05). Compared with the si-HCP5 group and si-HCP5+ anti-NC group, the si-HCP5+ anti-miR-525-5p group showed significant upregulation of cell apoptosis rate and levels of Bax and cleared Caspase-3 protein levels (all P<0.05), and significant downregulation of Bcl-2 protein levels ( P<0.05). Conclusion:lncRNA HCP5 may inhibit the expression of BTG2 by targeting up-regulation of miR-525-5p, thus leading to apoptosis of nerve cells in OGD/R models.
