Glutathione is a crucial guardian of protein integrity in the brain upon nitric oxide imbalance

Figures & data

Figure 1 Glutathione depletion is associated with 3-nitrotyrosine increase in mice brain without accumulation of apoptotic markers. (A) Left part: 20 µg of proteins extracted from mice brains were spotted on nitrocellulose membrane and subjected to Dot blot analysis using a polyclonal 3-nitrotyrosine (3-NT) antibody. Right part: density of immunoreactive dots was normalized for Ponceau Red spots and reported as arbitrary units (a.u.) and as means ± SD. (*p < 0.01 versus control, **p < 0.01 versus BSO-treated; n = 3). (B) 20 µg of total protein extracts were loaded for detection of Bcl-2, Bax and pro-caspase-3 by western blot. Actin was used as loading control. (C) mitochondria were purified from mice brains homogenates by Percoll^® gradient, lysed and 20 µg of proteins were loaded for detection of AIF by western blot. Cytochrome c was used as loading control.

Table 1 GSH concentration in mice treated with BSO

	Heart	Skeletal muscle	Lung	Liver	Brain
Control	4.17 ± 0.40	2.45 ± 0.71	9.43 ± 0.98	23.43 ± 4.23	5.25 ± 0.07
L-NAME	5.04 ± 1.05	2.00 ± 0.14	6.90 ± 0.04	25.01 ± 3.40	5.19 ± 0.13
BSO	0.44 ± 0.08**	0.20 ± 0.01*	3.15 ± 0.20**	14.32 ± 3.30*	4,50 ± 0.41*
L-NAME+BSO	0.49 ± 0.02**	0.19 ± 0.01*	3.10 ± 0.28**	6.95 ± 3.15*	4.60 ± 0.28*

C57/BL6 mice were treated with BSO (20 mM) and/or L-NAME (4 mM) in drinking water. After one month, GSH concentration was assayed on various homogenized organs and tissues by HPLC. Data are expressed as nmoles of GSH/mg of proteins and reported as means ± S.D. (*p<0.05; **p<0.001 versus control; n = 3).