A Plant-Derived Antioxidant Supplement Prevents the Loss of Retinal Ganglion Cells in the Retinas of NMDA-Injured Mice

Figures & data

Table 1 The Composition of the Oral Supplement. The Units are All in mg. PBS Was Used as Vehicle, and the Mice of Control Group Were Taken Same Volume of PBS

Structual Component	0.4 mg/ 200 µL	4 mg/ 200 µL
Hesperidin	0.02	0.2
Crocetin	0.004	0.04
Tamarindus indica	0.02	0.2
Reduced maltose starch syrup	0.272	2.72
Partly pregelatinized starch	0.08	0.8
Stearic acid calcium	0.004	0.04

Figure 1 Oral supplementation reduced lipid peroxidation after NMDA injury. The amount of MDA in the retina was evaluated with a TBARS assay 6 hours after the intravitreal injection of NMDA without supplementation or with a low- or high-dose supplement (16 mg/kg, 160 mg/kg). The average amount of MDA was calculated and is shown per mg of retinal protein (each group: n = 6). Data represent mean ± SD. ***p < 0.001.

Figure 2 Oral supplementation reduced the cleavage of α-fodrin in the retina after NMDA injury. (A) Immunoblot analysis of α-fodrin in retinas without supplementation or with a low- or high-dose supplement 6 hours after NMDA injury. Representative immunoreaction image with anti-α-fodrin showing intact α-fodrin (280 kDa) and calpain-cleaved fragmented α-fodrin (145 and 150 kDa). β-actin was used as an internal control. (B) The relative density of the cleaved-fodrin immunoreactive band. Relative density was based on the cleaved-fodrin immunoreactive band 6 hours after NMDA injection. Data represent mean ± SD (each group: n = 6). *p <0.05.

Figure 3 Decrease in TUNEL-positive cells after NMDA injury and supplementation. (A) Representative overlay photographs of retinal sections in mice with or without supplementation 24 hrs after NMDA injection. Red: TUNEL assay; blue: DAPI nuclear staining. Scale bar: 100 µm. (B) Histograms showing the TUNEL-positive cell count in the GCL of mice (non-supplementation group: n = 7, other groups: n = 8). Data represent mean ± SD, **P < 0.01, ***P < 0.001.

Abbreviations: GCL, ganglion cell layer; INL, inner nuclear layer.

Figure 4 Increase in RBPMS-positive cells after NMDA injury and supplementation. (A) Representative images of RBPMS-positive RGCs 24 hours after the intravitreal injection of NMDA without supplementation or with a low- or high-dose supplement. GCL, ganglion cell layer; INL, inner nuclear layer. Scale bar: 100 µm. (B) Histogram showing the average number of RBPMS-positive cells in each group. Data represent mean ± SD (PBS, n = 8; low-dose supplement, n = 6; high-dose supplement, n = 8). *p <0.05.