The effect of ethanol on erythrocyte carbonic anhydrase isoenzymes activity: An in vitro and in vivo study

Figures & data

Figure 1 SDS-polyacrylamide gel electrophoresis of HCA-I and HCA-II are purified by affinity gel (HCA: Human carbonic anhydrase, lane 1 is HCA-II, lane 2 is HCA-I).

Table I.  Purification scheme of HCA-I and HCA-II from human erythrocytes by Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography (HCA: Human carbonic anhydrase). Experimental analysis were performed at +4°C. CA enzyme activity was determined colorimetrically using CO₂-hydration method of Wilbur and Anderson. The enzyme unit (EU) was calculated by using the Equation (to-tc/tc) where to and tc are the times for pH change of the nonenzymatic and the enzymatic reactions, respectively.

Purification step	Activity (U/mL)	Total volume (mL)	Protein (mg/ml)	Total protein (mg)	Total Activity (U)	Spesific Activity (U/mg)	Yield (%)	Purification factor
Haemolysate	775	156	108	16848	120900	7.176	100	1
Sepharose-4B-L-tyrosine-sulfanilamide affinity chromatography	HCA-I	2932	23	0.68	15.64	67436	4311.8	55.78	600.9
HCA-II	3514	21	0.26	5.46	73794	13515.4	60.03	1883.4

Figure 2 Effect of the ethanol at different concentrations on human erythrocyte CA-I activity (CA-I: Carbonic anhydrase isoform-I).

Figure 3 Effect of the ethanol at different concentrations on human erythrocyte CA-II activity (CA-II: Carbonic anhydrase isoform-II).

Table II.  The in vivo effects of dosage of ethanol (2 mL.kg⁻¹ body weight) on rat red blood cell CA activity (CA: Carbonic anhydrase, n: 8).

Time after administration (h)	Total CA activity (%)^1,2
0	100.00 ± 4.13
1	85.29 ± 5.92*
3	66.60 ± 4.89**
5	92.56 ± 3.12

¹ Results are expressed as the Mean ± Standard Deviation, n = 8

² * p < 0.05, ** p < 0.01, vs control by Student's t test. Control value is enzyme activity before ethanol administration; 0 h.