Camp. Biochem. Physiol. Vol. 103C,No. 1, pp. 143-147, 1992 Printed in Great Britain
0
0306~4492/92 $5.00 + 0.00 1992 Pergamon Press Ltd
COMBINED EFFECT OF VANADIUM AND ZINC ON CERTAIN SELECTED HAEMATOLOGICAL INDICES IN RATS HALINA ZAPOROWSKAand WACLAW WASILEWSKI Department of Cell Biology, Institute of Biology, Maria Curie-SkIodowska University, 20-033 Lublin, Akademicka 19, Poland (Received 2 1 January 1992) Abstract-l. Two-month-old Wistar rats of both sexes received, as sole drinking liquid, an aqueous solution of ammonium metavanadate (AMV) and zinc chloride (ZC) at concentrations of 0.30 mg V/cm3 and 0.12 mg Zn/cm) respectively, for a period of 4 weeks. 2. The reference groups received for drinking at this time: water, AMV or ZC solutions at the same concentration. 3. In all groups of animals there was a statistically significant decrease in the uptake of food, AMV or ZC, as well AMV-ZC solutions, as compared with the food and water taken up by the control group. 4. In the group of animals receiving AMV or AMV-ZC solution for drinking the body weight increment diminished significantly. 5. In the animals drinking the AMV-ZC solution a statistically significant decrease in the erythrocyte count and haemoglobin level in the peripheral blood were recorded, similar to the groups drinking AMV or ZC solution. 6. In rats drinking aqueous AMV or ZC solutions and in females receiving AMV-ZC solution the percentage of recticulocytes and polychromatophilic erythrocytes increased, moreover, in the peripheral blood. It was not. however, associated with marked percentage changes in the composition of the bone marrow cells.
INTRODIJCI’ION Zinc is an essential trace element for animals and humans. Its physiological role and consequences of
its deficiency have been the subject of numerous experimental and review papers (Fisher, 1975; Kang et al., 1977; Shah et al., 1979; Johanning and O’Dell, 1989; Sato and Nagai, 1989; Smogorzewska, 1989). This element is present in many metaloenzymes, it affects the metabolism of nucleic acids, proteins, lipids and has a stabilising influence on the cell membranes (Chvapil, 1976; Prasad, 1979; Piechota, 1983; Bednarek, 1988). It may also play the role of an antioxidant (Bray and Bettger, 1990). It has, moreover, been demonstrated that zinc attenuates the toxic effects of such elements as lead (Flora and Tandon, 1990), mercury (Fukino et al., 1986), copper (Bremner et al., 1976) and of some compounds, for example ethanol (Dar et al., 1986; Seyoum and Persaud, 1991), carbon tetrachloride (Chvapil et al., 1973) and of bromobenzene (Szymanska et al., 1991). Zinc salts are also a new class of radioprotectors (Floersheim and Bieri, 1990). In excessive doses, however, zinc has a toxic influence (Fosmire, 1990). Only a single mention was found on the role of zinc in vanadium intoxications (Venugopal and Luckey, 1978; Yamaguchi et al., 1989). Therefore, the present study was aimed at investigating the effect of simultaneous oral administration of vanadium and zinc on certain haematological indices in Wistar rats. MATERIALS
AND METHODS
The experiments were performed on 2-month-old Wistar rats of both sexes (54 males and 64 females). Within each
sex the animals were divided randomly into four groups. Group I (control) received water to drink. The remaining animals were given solely the following solutions: group II-aqueous solution of ammonium metavanadateNH,VO1 (AMV; Reachim, U.S.S.R.) containing 0.30 mg V/ cm’; group III-aqueous solution of zinc chloride-ZnCl, (ZC; Fluka, Switzerland) of 0.12 mg Zn/cm’ concentration and group IV AMV-ZC aqueous solution of 0.30 mg V and 0.12 mg Zn/cm3 concentration. All the animals were fed a standard granulated rodent laboratory chow-LSM (CLPP, Motycz near Lublin). They were kept in cages singly under constant conditions (temperature 18-2o”C, relative air humidity 60 + lo%, at natural day-night light cycles). Every day the amount of food and the volume of liquid drunk by the tested groups of animals were controlled. After four weeks the rats were bled under ether anaesthesia and the erythrocyte and leukocyte counts, haemoglobin level and haematocrit were detemined by standard methods. In the peripheral blood smears stained by Pappenheim’s method the leukocyte picture was analysed and the percentage of polychromatophilic erythrocytes was determined. On bone marrow smears stained by the same method the percentage of nuclear blood cells was estimated. Moreover, the percentage of reticulocytes was determined in peripheral blood preparations stained with Brilliant Cresyl Blue. The results were subjected to statistical analysis by Students f-test at the significance level P < 0.05. RESULTS In rats receiving an aqueous ammonium metavanadate solution (AMV) as sole drinking liquid or an
ammonium metavanadate and zinc chloride solution (AMV-ZC) for a period of four weeks diarrhoea was frequently observed. In five cases in males and seven in females it ended in death. In both discussed animal groups a statistically significant decrease in the body 143
HALINA ZAPOROWSKAand WAUAW
144
L
1
I
I
I
I
0
1
2
3
L
weeks
Fig. 1. Body weight increment of Wistar rats in chronic ammonium metavanadate (AMV) or/and zinc chloride (ZC) intoxication. The graph shows: A-_-A control group; A--A group intoxicated with ZC; n -m group intoxicated with AMV; 0-O group intoxicated with AMV-ZC. Points represent mean values +S.E.M. increment was also observed as compared with that in the controls. The lowest body weight
weight
Table I. The effect of ammonium
metavanadate
WASLEWSKI
increment was noted in the group receiving the aqueous solution of AMV-ZC. In some of the individuals in this group even a decrease in body weight was observed. On the other hand, the body weight increment of the animals given the aqueous solution of zinc chloride (ZC) was similar to that of the controls (Table 1, Fig. 1). In all the three groups of animals a statistically significant decrease in the amount of food, AMV, ZC and AMV-ZC solutions consumed was noted as compared with food and water taken up by the control group (Table 2). A statistically significant decrease in the erythrocyte count and haemoglobin level was found in the peripheral blood of these animals. The haematocrit index fell only slightly (Table 3). The percentage of reticulocytes and polychromatophilic erythrocytes in the peripheral blood of these animals increased significantly as compared with that in the controls. Only in the males drinking the AMV-ZC solution the values of these parameters were similar to those of the control ones (Fig. 2). Moreover, in rats receiving ZC or AMV-ZC solution an increase in the leukocyte count was noted, the difference being statistically significant only in the group of males drinking the ZC solution (Table 3). The rise in the leukocyte count resulted above all from an increased lymphocyte number (Table 4). Analysis of the bone marrow showed no major changes in the percentage composition of cells between the control and the groups of rats receiving ZC or the AMV-ZC solution (Table 5). DISCUSSION
In their (Zaporowska
(AMV) or/and
zinc chloride
earlier investigations the authors and Wasilewski, 1989) found that in (ZC) on body weight gain in tested rats Body weight gain
Group of animals
Initial number of animals
Mortality n
Number of animals examined
g/24 hr
g
Males Control AMV zc AMV-ZC
12 13 13 16
0 2 0 3
12 II 13 13
84.83 39.68 78.81 17.50
f + f f
Females Control AMV zc AMV-ZC
12 14 17 21
0 3 0 4
12 II 17 17
36.70 22.73 40.53 10.12
_+2.41 + S.91* f 2.49 _+ 3.73’1.
The results are given as mean values f S.E.M. Significantly different from control group: *P c 0.05; l**P
Table 2. Food, fluid, vanadium Group of animals
Food intake g/rat/24 hr
0
0306~4492/92 $5.00 + 0.00 1992 Pergamon Press Ltd
COMBINED EFFECT OF VANADIUM AND ZINC ON CERTAIN SELECTED HAEMATOLOGICAL INDICES IN RATS HALINA ZAPOROWSKAand WACLAW WASILEWSKI Department of Cell Biology, Institute of Biology, Maria Curie-SkIodowska University, 20-033 Lublin, Akademicka 19, Poland (Received 2 1 January 1992) Abstract-l. Two-month-old Wistar rats of both sexes received, as sole drinking liquid, an aqueous solution of ammonium metavanadate (AMV) and zinc chloride (ZC) at concentrations of 0.30 mg V/cm3 and 0.12 mg Zn/cm) respectively, for a period of 4 weeks. 2. The reference groups received for drinking at this time: water, AMV or ZC solutions at the same concentration. 3. In all groups of animals there was a statistically significant decrease in the uptake of food, AMV or ZC, as well AMV-ZC solutions, as compared with the food and water taken up by the control group. 4. In the group of animals receiving AMV or AMV-ZC solution for drinking the body weight increment diminished significantly. 5. In the animals drinking the AMV-ZC solution a statistically significant decrease in the erythrocyte count and haemoglobin level in the peripheral blood were recorded, similar to the groups drinking AMV or ZC solution. 6. In rats drinking aqueous AMV or ZC solutions and in females receiving AMV-ZC solution the percentage of recticulocytes and polychromatophilic erythrocytes increased, moreover, in the peripheral blood. It was not. however, associated with marked percentage changes in the composition of the bone marrow cells.
INTRODIJCI’ION Zinc is an essential trace element for animals and humans. Its physiological role and consequences of
its deficiency have been the subject of numerous experimental and review papers (Fisher, 1975; Kang et al., 1977; Shah et al., 1979; Johanning and O’Dell, 1989; Sato and Nagai, 1989; Smogorzewska, 1989). This element is present in many metaloenzymes, it affects the metabolism of nucleic acids, proteins, lipids and has a stabilising influence on the cell membranes (Chvapil, 1976; Prasad, 1979; Piechota, 1983; Bednarek, 1988). It may also play the role of an antioxidant (Bray and Bettger, 1990). It has, moreover, been demonstrated that zinc attenuates the toxic effects of such elements as lead (Flora and Tandon, 1990), mercury (Fukino et al., 1986), copper (Bremner et al., 1976) and of some compounds, for example ethanol (Dar et al., 1986; Seyoum and Persaud, 1991), carbon tetrachloride (Chvapil et al., 1973) and of bromobenzene (Szymanska et al., 1991). Zinc salts are also a new class of radioprotectors (Floersheim and Bieri, 1990). In excessive doses, however, zinc has a toxic influence (Fosmire, 1990). Only a single mention was found on the role of zinc in vanadium intoxications (Venugopal and Luckey, 1978; Yamaguchi et al., 1989). Therefore, the present study was aimed at investigating the effect of simultaneous oral administration of vanadium and zinc on certain haematological indices in Wistar rats. MATERIALS
AND METHODS
The experiments were performed on 2-month-old Wistar rats of both sexes (54 males and 64 females). Within each
sex the animals were divided randomly into four groups. Group I (control) received water to drink. The remaining animals were given solely the following solutions: group II-aqueous solution of ammonium metavanadateNH,VO1 (AMV; Reachim, U.S.S.R.) containing 0.30 mg V/ cm’; group III-aqueous solution of zinc chloride-ZnCl, (ZC; Fluka, Switzerland) of 0.12 mg Zn/cm’ concentration and group IV AMV-ZC aqueous solution of 0.30 mg V and 0.12 mg Zn/cm3 concentration. All the animals were fed a standard granulated rodent laboratory chow-LSM (CLPP, Motycz near Lublin). They were kept in cages singly under constant conditions (temperature 18-2o”C, relative air humidity 60 + lo%, at natural day-night light cycles). Every day the amount of food and the volume of liquid drunk by the tested groups of animals were controlled. After four weeks the rats were bled under ether anaesthesia and the erythrocyte and leukocyte counts, haemoglobin level and haematocrit were detemined by standard methods. In the peripheral blood smears stained by Pappenheim’s method the leukocyte picture was analysed and the percentage of polychromatophilic erythrocytes was determined. On bone marrow smears stained by the same method the percentage of nuclear blood cells was estimated. Moreover, the percentage of reticulocytes was determined in peripheral blood preparations stained with Brilliant Cresyl Blue. The results were subjected to statistical analysis by Students f-test at the significance level P < 0.05. RESULTS In rats receiving an aqueous ammonium metavanadate solution (AMV) as sole drinking liquid or an
ammonium metavanadate and zinc chloride solution (AMV-ZC) for a period of four weeks diarrhoea was frequently observed. In five cases in males and seven in females it ended in death. In both discussed animal groups a statistically significant decrease in the body 143
HALINA ZAPOROWSKAand WAUAW
144
L
1
I
I
I
I
0
1
2
3
L
weeks
Fig. 1. Body weight increment of Wistar rats in chronic ammonium metavanadate (AMV) or/and zinc chloride (ZC) intoxication. The graph shows: A-_-A control group; A--A group intoxicated with ZC; n -m group intoxicated with AMV; 0-O group intoxicated with AMV-ZC. Points represent mean values +S.E.M. increment was also observed as compared with that in the controls. The lowest body weight
weight
Table I. The effect of ammonium
metavanadate
WASLEWSKI
increment was noted in the group receiving the aqueous solution of AMV-ZC. In some of the individuals in this group even a decrease in body weight was observed. On the other hand, the body weight increment of the animals given the aqueous solution of zinc chloride (ZC) was similar to that of the controls (Table 1, Fig. 1). In all the three groups of animals a statistically significant decrease in the amount of food, AMV, ZC and AMV-ZC solutions consumed was noted as compared with food and water taken up by the control group (Table 2). A statistically significant decrease in the erythrocyte count and haemoglobin level was found in the peripheral blood of these animals. The haematocrit index fell only slightly (Table 3). The percentage of reticulocytes and polychromatophilic erythrocytes in the peripheral blood of these animals increased significantly as compared with that in the controls. Only in the males drinking the AMV-ZC solution the values of these parameters were similar to those of the control ones (Fig. 2). Moreover, in rats receiving ZC or AMV-ZC solution an increase in the leukocyte count was noted, the difference being statistically significant only in the group of males drinking the ZC solution (Table 3). The rise in the leukocyte count resulted above all from an increased lymphocyte number (Table 4). Analysis of the bone marrow showed no major changes in the percentage composition of cells between the control and the groups of rats receiving ZC or the AMV-ZC solution (Table 5). DISCUSSION
In their (Zaporowska
(AMV) or/and
zinc chloride
earlier investigations the authors and Wasilewski, 1989) found that in (ZC) on body weight gain in tested rats Body weight gain
Group of animals
Initial number of animals
Mortality n
Number of animals examined
g/24 hr
g
Males Control AMV zc AMV-ZC
12 13 13 16
0 2 0 3
12 II 13 13
84.83 39.68 78.81 17.50
f + f f
Females Control AMV zc AMV-ZC
12 14 17 21
0 3 0 4
12 II 17 17
36.70 22.73 40.53 10.12
_+2.41 + S.91* f 2.49 _+ 3.73’1.
The results are given as mean values f S.E.M. Significantly different from control group: *P c 0.05; l**P
Table 2. Food, fluid, vanadium Group of animals
Food intake g/rat/24 hr
