Suppression of glutathione system and upregulation of caspase 3-dependent apoptosis mediate rohypnol-induced gastric injury

Figures & data

Table 1. Effect of rohypnol on body weight gain and relative gastric weight.

Group/parameter	Control	Low-dose rohypnol	High-dose rohypnol	Low dose- rohypnol recovery	High dose- rohypnol recovery
Body weight gain (g)	46.50 ± 3.52	48.25 ± 3.61	44.50 ± 3.34	51.00 ± 3.50	47.50 ± 3.14
Absolute stomach weight (g)	14.6 ± 1.80	12.7 ± 1.33	11.7±1.73*	14.8 ± 1.80^∼	13.4 ± 1.93
Relative stomach weight (%)	6.15 ± 0.76	5.34 ± 0.880	4.95 ± 0.69*	6.11 ± 0.89	5.39 ± 0.92

Notes: Values are presented as mean ± SD. *P < .05 vs control; ^#P < .05 vs low-dose; ^∼P < .05 vs high-dose.

Figure 1. Effect of rohypnol on gastric levels of malondialdehyde, MDA (A), reduced glutathione, GSH (B), oxidized glutathione (C), and GSH/GSSG ratio (D). Values are presented as mean ± SD. *P < .05 vs control; #P < .05 vs low-dose; ∼P < .05 vs high-dose, ^P < .05 vs low-dose recovery.

Table 2. Effect of rohypnol on gastric enzymatic antioxidant activities.

Group/parameter	Control	Low-dose rohypnol	High-dose rohypnol	Low dose-rohypnol recovery	High dose-rohypnol recovery
Superoxide dismutase (U/mg)	156.6 ± 16.97	119.9 ± 9.63*	100.5 ± 13.5*	138.3 ± 11.2^∼	125 ± 21.6*^∼
Catalase (μmol/g)	6.04 ± 0.53	5.28 ± 0.67	4.27 ± 1.12*	4.96 ± 1.13	3.46 ± 1.15*^#
Glutathione-S-transferase (U/mg)	5.53 ± 0.79	4 ± 0.55*	2.66 ± 0.35*#	4.16 ± 1.1*^∼	3.2 ± 1.0*
Glutathione peroxidase (μmol/g)	6.51 ± 0.82	3.81 ± 0.76*	2.83 ± 0.60*	3.59 ± 1.43*	2.53 ± 0.95*

Notes: Values are presented as mean ± SD. *P < .05 vs control; ^#P < .05 vs low-dose; ^∼P < .05 vs high-dose.

Figure 2. Effect of rohypnol on gastric levels of nitric oxide, NO, (A), tumor necrotic factor-α, TNF-α (B), and interleukin-6, IL-6 (IL-6) (C). Values are presented as mean ± SD. *P < .05 vs control; #P < .05 vs low-dose; ∼P < .05 vs high-dose, ^P < .05 vs low-dose recovery.

Figure 3. Effect of Rohypnol on gastric levels of cyclo-oxygenase-2 (COX-2). Values are presented as mean ± SD. *P < .05 vs control; #P < .05 vs low-dose; ∼P < .05 vs high-dose.

Table 3. Effect of rohypnol on gastric genotoxicity and apoptotic markers.

Group/parameter	Control	Low-dose rohypnol	High-dose rohypnol	Low dose- rohypnol recovery	High dose- rohypnol recovery
8-OHdG (ng/mg)	0.49 ± 0.07	0.68 ± 0.07*	0.77 ± 0.12*	0.59 ± 0.37	0.69 ± 0.16*
Gastric DFI (%)	17 ± 3.30	36 ± 4.04*	47.75 ± 5.4*#	29.63 ± 8.2*^∼	45.13 ± 2.95*#
Caspase-3 (ng/mg)	0.368 ± 0.22	0.561 ± 0.18	0.86 ± 0.09*	0.43 ± 0.26^∼	0.77 ± 0.37*

Notes: Values are presented as mean ± SD. *P < .05 vs control; #P < .05 vs low-dose; ^∼P < .05 vs high-dose.

Figure 4. Photomicrographs of the gastric tissue. The gastric tissues of the control (A), low dose Rohypnol-treated (B), and the low dose recovery (D) and high dose recovery (E) groups showed preserved layers; the mucosa (line) lined by simple columnar epithelium lying on a thin layer of the muscularis mucosa (MM, broken black arrow), and the submucosa (SM in red circle) composed of branches of blood vessels (white arrow in blue circle) and lymphatics, the muscularis (M in black circle) and adventitia. The gastric glands (GG in green circle) and gastric pit (GP in yellow circle) appeared unremarkable. However, the gastric tissue of the high dose Rohypnol-treated (C) revealed hyperemic blood vessels (white arrow in blue circle) and moderately atrophied gastric gland (GG in green circle).

Table 4. Effect of rohypnol on gastric mucosa integrity.

Group/parameter	Control	Low-dose rohypnol	High-dose rohypnol	Low dose- rohypnol recovery	High dose- rohypnol recovery
Gastric mucosa lesion index	0.20 ± 0.44	0.60 ± 0.54	2.20 ± 0.45*^#	0.40 ± 0.55^∼	0.80 ± 0.83^∼
Gastric mucosa pathological injury integral	0.25 ± 0.50	0.75 ± 0.50	3.00 ± 0.81*^#	0.50 ± 0.57^∼	0.75 ± 0.50^∼

Notes: Values are presented as mean ± SD. *P < .05 vs control; #P < .05 vs low-dose; ^∼P < 0.05 vs high-dose.

Data availability statement

The data that support the findings of this study are available on request from the corresponding author.