Deranged Bioenergetics and Defective Redox Capacity in T Lymphocytes and Neutrophils Are Related to Cellular Dysfunction and Increased Oxidative Stress in Patients with Active Systemic Lupus Erythematosus
Figure 4
Comparison of intracellular reduced-form glutathione (GSH) levels, enzyme activity, and gene expression of glutathione peroxidase (GSH-Px) in the plasma and different blood cell populations of normal individuals. (a) Intracellular GSH levels (μM/ cells/mL) in T, PMN, red blood cells, platelets, and plasma of normal individuals. (b) GSH-Px enzyme activity (mU/ cells/mL) in plasma and different blood cells of normal individuals. One milliunit (mU) of GSH-Px enzyme activity is the activity that catalyzes the oxidation of 1 nmol NADPH/mL/min. (c) Expression of GSH-Px mRNA in T and PMN of two normal individuals by RT-PCR, lane 1: G3PDH (452 bp, as internal control), lane 2: GSH-Px (354 bp). (d) A representative case demonstrating dose-response expression of GSH-Px isomers in a normal T and a normal PMN by Western blot. Three doses (2 μL in lanes 1 and 4; 4 μL in lanes 2 and 5; 10 μL in lanes 3 and 6) of cell lysates (protein concentration 26 mg/mL) were analyzed in Western blot probed by antihuman GSH-Px antibody. Four GSH-Px isomers are identified as monomer (25 kDa), dimer (50 kDa), trimer (75 kDa), and tetramer (100 kDa). Two normal samples exhibited a similar tendency.
