Effects of DJ-1 protein on retinal ganglion cells and visual function in mice after optic nerve crush injury
10.3760/cma.j.cn511434-20201218-00621
- VernacularTitle:DJ-1蛋白对视神经钳夹伤后小鼠视网膜神经节细胞及视功能的影响
- Author:
Lingyi OUYANG
;
Tao HE
;
Yiqiao XING
- From:
Chinese Journal of Ocular Fundus Diseases
2021;37(5):377-384
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of DJ-1 encoded by Park7 gene on retinal ganglion cells(RGC) and visual function after optic nerve crush injury (ONC) in mice.Methods:Thirty-seven and 116 healthy male C57BL/6J mice were randomly divided into group normal, group ONC 2d, group ONC 5d, group ONC 7d and group control, group Park7, group Park7-ONC, group ONC and group green fluorescent protein (GFP)-ONC. Group ONC 2d, group ONC 5d and group ONC 7d were sacrificed on the 2nd, 5th and 7th day after the establishment of ONC model, and the follow-up experiments were carried out. The mice in group Park7 and group Park7-ONC were injected 1 μ recombinant adeno-associated virus (rAAV) with knocking down Park7 gene into vitreous cavity, and 1 μl rAAV with only GFP was injected into vitreous cavity of mice in group GFP-ONC, and virus transfection was observed 4 weeks after injection. The injury of ONC was perfomed at 23 days after vitreous injection in group ONC, group Park7-ONC and group GFP-ONC, and the samples were taken for follow-up experiment 5 days after modeling. The average density of RGC was observed by immunofluorescence staining, the latencies and amplitudes of a-wave, b-wave and photopic negative response (phNR) and the amplitude of oscillatory potential (OPs)were detected by full-field flash electroretinogram,and the visual acuity of mice was measured by optomotor response (OMR). The relative expression levels of DJ-1, Bax and B lymphoblastoma / leukemia-2 (Bcl-2) protein in the retina of mice in each group were detected by Western blot. One-way ANOVA was used to compare the data between groups, and t-test was used for pairwise comparison between groups.Results:Compared with the normal group, the relative expression of DJ-1 protein in the retina of the ONC 2 d group and ONC 5 d group increased significantly, and the difference was statistically significant ( t=16.610, 5.628, P<0.01,<0.05). Four weeks after virus transfection, strong GFP expression was seen in the RGC layer and inner plexiform layer of the retina of mice in the Park7 group. Compared with the control group, the RGC density of the retina in the ONC group decreased significantly, and the difference was statistically significant ( t=16.520, P<0.000); compared with the ONC group, the RGC density of the retina in the Park7- ONC group decreased significantly, and the difference was statistically significant ( t=6.074, P<0.01). With the increase of stimulus light intensity, the dark adaptation a wave and b wave latency of the mice in the control group gradually shortened, and the amplitude gradually increased. The stimulus light intensity was 3 cd·s/m 2. There was no statistically significant difference in the dark adaptation a wave and b wave latency and amplitude of the control group, Park7 group, Park7-ONC group, ONC group, and GFP-ONC group (Incubation period: F=0.503, 2.592; P=0.734, 0.068. Amplitude: F=0.439, 1.451; P=0.779, 0.247). Compared with the control group, the Ops and PhNR amplitudes of the ONC group mice were significantly decreased ( t=15.07, 12.80; P<0.000,<0.001). Compared with the ONC group, the Ops and PhNR amplitudes of the mice in the Park7- ONC group were significantly decreased ( t=4.042, 5.062; P<0.05,<0.01); there was no statistically significant difference in the PhNR latency of the mice in each group ( F=1.327, P=0.287). Compared with the control group, the visual acuity of the mice in the ONC group was significantly decreased, and the difference was statistically significant ( t=23.020, P<0.000); compared with the ONC group, the visual acuity of the mice in the Park7-ONC group was significantly decreased, and the difference was statistically significant ( t=3.669, P<0.05). Compared with the control group, Park7-ONC group and ONC group, the relative expression of DJ-1 protein in the mouse retina was significantly down-regulated, and the difference was statistically significant ( t=47.140, 26.920; P<0.000,<0.000). There was no significant difference between ONC group and GFP-ONC group ( t=0.739, P=0.983). Compared with the ONC group, the relative expression of Bax protein in the mouse retina of the Park7-ONC group was significantly increased, and the relative expression of Bcl-2 protein was significantly reduced. The differences were statistically significant ( t=5.960, 9.710; P<0.05,<0.05); the relative expression ratio of Bcl-2/Bax in the Park7-ONC group was significantly lower than that in the ONC group, and the difference was statistically significant ( t=13.620, P<0.01). Conclusion:The expression of DJ-1 encoded by Park7 gene is down-regulated after Park7 gene was knocked down, which aggravates the RGC damage and the decrease of retinal electrophysiological response and visual function in ONC injury mice.
