Effects of some analgesic anaesthetic drugs on human erythrocyte glutathione reductase: an in vitro study

Figures & data

Table I..  Purification scheme of GR from human erythrocyte.

Purification step	Total volume (mL)	Activity (U/mL)	Total activity (U)	Protein (mg/mL)	Specific activity (U/mg)	Yield (%)	Purification factor
Hemolysate	165	0.33	54.45	34	0.0097	100	1
Ammonnium sulfate precipitation (30–70%)	43	0.77	33.11	37	0.021	60.81	2.16
2′, 5′-ADP Sepharose 4B affinity chromatography	21	1.57	32.97	0.41	3.83	60.55	395
Sephadex G-200 gel filtration chromatography	19	0.83	15.77	0.04	20.75	29	2139

Figure 1.  SDS-PAGE bands of GR (Lane 1: Standards: rabbit phosphorylase B (Mr 97.400), bovine albumin (66.000), chicken ovalbumin (45.000), and bovine carbonic anhydrase (29.000); Lane 2-3: Gel filtration chromatography.

Table II. K_i and IC₅₀ values obtained from regression analysis graphs for GR in the presence of different drugs concentrations.

Drugs	I50 (mM)	Ki (mM)	Inhibition type
Morphine	0.0128	0.0165 ± 0.0013	competitive
Etomidate	0.0523	0.0725 ± 0.0029	competitive
Tenoxicam	0.082	0.13 ± 0.025	competitive
Propofol	0.242	0.418 ± 0.056	Non-competitive
Lornoxicam	0.300	0.42 ± 0.18	competitive
Ketoprofen	6.278	22.14 ± 7.6	competitive
Diclofenac	7.265	23.97 ± 2.1	competitive

Figure 2.  Activity % vs [morphine] regression analysis graphs for human erythrocyte GR in the presence of 5 different morphine concentrations.

Figure 3.  Lineweaver-Burk graph for 5 different substrate (GSSG) concentrations and 3 different morphine concentrations for determination of K_i.