Fig 1.
Short-term hypoglycemic coma model.
(A) Protocol for hypoglycemic loading and induction of the hyperglycemic state. Rapid-acting insulin was intraperitoneally administered at a dose of 15 IU/kg. After 2 or 10 min of the presence of an isoelectric electroencephalogram (EEG), 50% glucose was intravenously injected at a dose of 0.2 mL and intravenously infused at a rate of 2.2 mL/h. Rats were sacrificed after 24 h of glucose reperfusion. (B) Alda-1 treatment protocol. After 2 min of an isoelectric EEG state, Alda-1 was administered intravenously at a dose of 8.5 mg/kg with 50% glucose (Alda-1 group) or the same dose of dimethyl sulfoxide (DMSO) was administered (control group).
Fig 2.
Temporal changes in blood glucose levels after administration of insulin and glucose.
Blood glucose levels were measured every 15 min before and after injection of rapid-acting insulin (from -15 min to 120 min; n = 48) as well as after injection of 50% glucose (from 15 min to 180 min and 24 h; n = 9).
Fig 3.
Temporal changes in electroencephalogram findings and neurological signs.
EEG; Electroencephalogram, N/A; not applicable. Data are means ± SD. Since pupil diameter and light reflexes were not maintained at a constant level during each phase, these findings were not suitable for predicting precise BG levels.
Fig 4.
Distribution of 4-HNE-positive cells in the short-term hypoglycemic coma model.
(A) Representative images of 4-HNE-positive cells in the sham group, the 2-min isoelectric EEG (iso-EEG) group, and the 10-min isoelectric EEG group. Higher magnification views of 4-HNE-positive cells are shown in the small frames. Scale bar: 100 μm. (B) The number of 4-HNE-positive cells in the sham group, the 2-min isoelectric EEG group, and the 10-min isoelectric EEG group. Data are means ± SD (n = 15 fields). *P < 0.001.
Fig 5.
Fluoro-Jade B-positive cells in the short-term hypoglycemic coma model.
(A) Representative images of Fluoro-Jade B-positive cells in the sham group, the 2-min isoelectric EEG (iso-EEG) group, and the 10-min isoelectric EEG group. Scale bar: 100 μm. (B) The number of Fluoro-Jade B-positive cells in the sham group, the 2-min isoelectric EEG group, and the 10-min isoelectric EEG group. Data are means ± SD (n = 15 fields). *P < 0.001.
Fig 6.
Effects of Alda-1 on 4-HNE-positive cells.
(A) Representative images of 4-HNE-positive cells treated with vehicle (dimethyl sulfoxide; DMSO) or Alda-1. A higher magnification view of a 4-HNE-positive cell is shown in the small frame. Scale bar: 100 μm. (B) Number of 4-HNE-positive cells in groups treated with vehicle (DMSO) or Alda-1. Data are means ± SD (n = 15 fields). *P = 0.002.
Fig 7.
Effects of Alda-1 on Fluoro-Jade B-positive cells.
(A) Representative images of Fluoro-Jade B-positive cells treated with vehicle (DMSO) or Alda-1. Scale bar: 100 μm. (B) Number of Fluoro-Jade B-positive cells in groups treated with vehicle (DMSO) or Alda-1. Data are means ± SD (n = 15 fields). *P = 0.020.