| Objective:Lead (Pb) and its compound are widely used in industrial and daily lives. Lead is a known chronic neurotoxicant. There is evidence that lead has been linked to neurodegenerative disorders. Employees in lead refinery could obviously be caused mental and hematological dysfunction. In the following experiments, lead-excretion glycan is used as a new type of lead excretion-food in mice. We investigate the potential genetoxic and reproductive poison of Lead-excretion Glycan and its effectiveness of lead excretion to evaluate its feasibility for use. At the same time, we would observe if Lead-excretion Glycan can cause the removal or replacement of other essential metals to provide foundation for seeking a safe and efficacious health food for lead overload.Methods1. MTD of Lead-excretion GlycanHealth mice of Kunming strain (7 males and 7 females) were given the Lead-excretion Glycan solution with the maximum concentration by intragastric administration method and observed for a week. The number of dead mice and the weight everyday were recorded.2. Peripheral blood and bone marrow cell micronucleus experiment of Lead-excretion Glycan30 healthy mice of Kunming strain (15 males and 15 females) were randomly divided into 5 groups according to their body weight: Cyclophosphamide positive control group, negative control group, high dose group (1/2MTD), medium dose group (1/4MTD), low dose group (1/8MTD), then peripheral blood and bone marrow cell micronucleus experiment of Lead-excretion Glycan was carried out 24 hours after the fourth application of medicine. The number of dead mice and the weight of mice everyday were recorded. 3. Sperm aberration experiment of Lead-excretion Glycan30 healthy male mice of Kunming strain were randomly divided into 5 groups according to their body weight: Cyclophosphamide positive control group, negative control group, high dose group(1/2MTD), medium dose group(1/4MTD), low dose group(1/8 MTD), the drug was applied for 5 successive days. On the 35th day after the 1st application of Lead-excretion Glycan, sperm aberration experiment was performed. The number of dead mice and the weight of mice everyday were recorded.4. Effect of Lead-excretion Glycan removing lead50 healthy mice of Kunming strain (25 males and 25 females) were randomly divided into 5 groups according to their body weight: negative control group, high-dose group, medium-dose group, low-dose group, model group. Pb(Ac)2 solution were given to the 5 groups except the negative control group to form lead-poisoned model in mice, then low-dose group, medium-dose group and high-dose group were given various dose of Lead-excretion Glycan. The negative control group and the model group were given normal saline. The mice were sacrificed by decapitation after the last administration and then the blood, liver, kidney and brain tissues were removed. The contents of lead in blood, liver, kidney, bone and brain were determined by graphite furnace atomic absorption spectrophotometer. The number of dead mice and the weight of mice every 5 days were recorded.5. Effects of Lead-excretion Glycan removing other essential metal elementsThe content of calcium, magnesium, iron, zinc, copper and manganese contents in the blood, liver, kidney, bone and brain tissues of the mice were determined by the flame atomic absorption spectrophotometer.6. Inhibition of lead-excretion glycan on micronuclei formation induced by lead acetate in mice6.1 Peripheral blood and bone marrow cell microcleus experiment120 healthy mice of Kunming strain (60 males and 60 females) were randomly divided into 6 groups according to their body weight: Cyclophosphamide positive control group, negative control group, lead-exposed group, high-dose group, medium-dose group and low-dose group. The mice of various dose groups treated with lead acetate were administered different dose of the lead-excretion glycan. Peripheral blood and bone marrow cell micronucleus experiment 48 hours after the 2nd and 4th application of lead-excretion glycan. The number of dead mice and the Weight of mice everyday were recorded. 6.2 Effect of Lead-excretion Glycan removing lead from liver, kidney, and brain in mice suffering acute intoxation of lead60 healthy mice of Kunming strain (30 males and 30 females) were randomly divided into 6 groups according to their body weight: Cyclophosphamide positive control group, negative control group, lead-exposed group, high-dose group, medium-dose group, low-dose and medium-dose preventive group. The mice were sacrificed by decapitation after 4th administration. The selected organs (liver, kidney, brain) were removed, weighted and frozen for later analysis of lead by graphite atomic absorption spectrophotometer.6.3 Effect of Lead-excretion Glycan removing other essential metal elements such as calcium, magnesium, iron, zinc, copper in mice suffering acute intoxation.60 healthy mice of Kunming strain (30 males and 30 females) were randomly divided into 6 groups according to their body weight: Cyclophosphamide positive control group, negative control group, high-dose group, medium-dose group, low-dose and medium-dose preventive group. The mice were sacrificed by decapitation after 4th administration. The selected organs (liver, kidney, brain) were removed, weighted and frozen for later analysis of calcium, magnesium, iron, zinc, copper by the flame atomic absorption spectrophotometer.7. Effect of Lead-excretion glycan recovering sperm abnormalities induced by lead acetate in mice49 healthy male mice of Kunming strain were randomly divided into 7 groups according to their weight: Cyclophosphamide positive control group, negative control group, lead-exposed group, high-dose group, medium-dose group, low-dose group, medium-dose preventive group. Lead-excretion glycan was applied for 5 successive days. On the 35th day after the 1st application of medicine, sperm aberration experiment was carried out. The number of dead mice and the weight of mice everyday were recorded. All kinds of sperm abnormality was observed under high power lens and classified into normal and different abnormal types as hookless, banana, amorphous, double heads, double tails, balloons.8. Effect of recovering the anti-oxidizing system of Lead-excretion Glycan36 healthy mice of Kunming strain (18 males and 18 females) were randomly divided into 6 groups according to their body weight: model group, Cyclophosphamide positive control group, negative control group, high-dose group, medium-dose group, low-dose and medium-dose preventive group. Pb(Ac)2 solution was given to the 6 groups except the negative control group to make lead-poisoned model in mice, then different dose of lead-excretion glycan was administered except model group. Lead-excretion Glycan was given to the preventive group while Pb(Ac)2 solution was applied. The activities of SOD, GSH-PX and the content of MDA in the homogenate of liver, kidney and brain tissues of the mice were determined in every group. The number of dead mice and the weight of mice everyday were recorded during the experiment.The liver, kidney and brain of the mice were fixed with 40% paraformaldehyde, and then hematoxylin-eosin stained. Pathomorphological observations were carried out to investigate the morphological changes.9. Effect of recovering hepatic function and renal function of Lead-excretion glycan against lead in mice.36 healthy mice of Kunming strain (18 males and 18 females) were randomly divided into 6 groups according to their body weight: model group, Cyclophosphamide positive control group, negative control group, high-dose group, medium-dose group, low-dose and medium-dose preventive group. Pb(Ac)2 solution was given to the 6 groups except the negative control group to make lead-poisoned model in mice. Then different dose of lead-excretion glycan was administrated except model group. Lead-excretion Glycan was given to the preventive group while Pb(Ac)2 solution was applied. Total proteins, albumin content, the activity of ALT, AST and ALP in serum were measured separately to evaluate hepatic function in every group;The content of blood uric acid (BUA) and content of the Blood urea nitrogen (BUN) were measured to evaluate renal function in every group.Results1. MTD of Lead-excretion GlycanThe results showed that the MTD of Lead-excretion Glycan is larger than 2.4g/kg; Lead-excretion Glycan has no effect on weight and no damage on liver, kidney and brain in mice.2. Peripheral blood and bone marrow cell micronucleus experiment of Lead-excretion GlycanThe results showed that there were no significant difference of the PCE micronucleus rate of the peripheral blood and bone marrow cell in high-dose group, medium-dose group, low-dose group in comparison with the negative control group (P>0.05), while the rate of the peripheral blood and bone marrow cell in CP group was higher than that of other groups (P<0.05).3. Sperm aberration experiment of Lead-excretion GlycanThe results showed that there were no significant differences of sperm aberration rate in high-dose group, medium-dose group and low-dose group in observations (P>0.05), while sperm aberration rate of CP positive control group was higher than that of all other groups(P<0.05).4. Effect of Lead-excretion Glycan removing leadThe contents of lead in the blood, liver, kidney, bone and brain in the treated groups were all lower than that of the model group (P<0.05) without dose response relation, but higher than that of negative group (P<0.05).5. Effects of lead-excretion Glycan removing other essential metal elementsNo significant difference existed among various groups in the contents of calcium in blood (P>0.05). In the content of calcium in liver and kidney, negative control group is higher than low-dose group and model group (P<0.05), but no significant difference between medium-dose group and high-dose group (P>0.05). The content of calcium in liver and kidney of model group was lower than that of negative group, medium-dose group and high-dose group (P<0.05). The content of calcium in bone of negative group was higher than that of other groups (P<0.05). The content of calcium in brain of model group was lower than that of negative group and high, medium, low dose groups, but no significant difference among negative group and high, medium, low dose groups(P>0.05). The content of calcium in brain of low-dose group and medium-dose group was lower than that of high-dose group (P<0.05).Magnesium content of high-dose group in blood is higher than that of negative control group (P<0.05). Magnesium content in bone of model group and high-dose group was lower than that of negative group, low-dose and medium dose group (P<0.05). No significant differences existed among low, medium dose group and negative control group(P>0.05). Magnesium content of model group and high-dose group in bone is lower than that of negative control group, low-dose group and medium-dose group (P<0.05). As far as magnesium content is concerned, no significant differences existed between high-dose group and model group (P>0.05). In comparison with magnesium content in liver, brain and kidney, no significant differences existed among low, medium dose group and negative control group(P>0.05). Iron content of negative group in livers was higher than that of low-dose group and model group (P<0.05). Iron content of medium-dose group and high-dose group is higher than that of model group; there were no significant differences among low, medium, high dose groups (P>0.05). In blood, bones, kidneys and brains, no significant differences existed among various groups (P>0.05).There were no significant differences among various groups in the contents of zinc in blood, livers and kidneys (P>0.05). In bones zinc content of negative control group and low dose, medium-dose, high-dose groups were higher than that of model group (P<0.05). As far as zinc content is concerned, no significant differences existed among low-dose, medium-dose high-dose groups and negative group (P<0.05).No significant differences existed among all groups in the content of copper and manganese in blood, liver, kidney, bone and brain (P>0.05). 6. Inhibition of lead-excretion glycan on micronuclei formation induced by lead acetate in mice6.1 Peripheral blood and bone marrow cell micronucleus experimentThe results after 2nd administration showed that the micronuclei rate in peripheral blood and bone marrow of mice in low, medium and high dose groups was significantly lower than that of the CP positive group(P<0.01) and lead-exposed group(P<0.05), higher than the negative control(P0.05). There were significant difference in micronuclei rate of peripheral blood and bone marrow of low group in comparison with high-dose group and medium-dose group (P<0.05). There were no significant differences in micronuclei rate of peripheral blood and bone marrow among medium-dose and low-dose group (P>0.05).The results after 4th administration showed that the micronuclei rate in peripheral blood and bone marrow of mice in low, medium and high dose groups was significantly lower than that of CP positive group(P<0.01) and model group(P<0.05), higher than that of the negative control(P<0.05). There were significant differences of micronuclei rate in peripheral blood and bone marrow of low, medium groups in comparison with high dose groups (P<0.05). There were no significant differences in micronuclei rate of peripheral blood and bone marrow among medium-dose and high-dose group (P>0.05).6.2 Effect of Lead-excretion Glycan removing lead from liver, kidney, and brain in mice suffering acute intoxationThe results showed that the contents of lead in the liver, brain and kidney in the Lead-excretion Glycan-treated groups were all lower than that of the lead-exposed group (P<0.05), higher than that of negative group (P<0.05).6.3 Effect of Lead-excretion Glycan removing other essential metal elements such . as calcium, magnesium, iron, zinc, copper in mice suffering acute intoxation.In comparison with calcium, magnesium, copper contents, no significant differences existed among various groups in the livers, kidneys, brains (P>0.05). Iron and zinc contents of Lead-excretion Glycan-treated groups in the livers and kidneys was higher than that of the lead-exposed group (P<0.05).7. Lead-excretion Glycan protect against the reproductive toxicity caused by leadCompared with the negative group, the sperm aberration rate of other groups was significantly higher (P<0.05) , and treated groups with high, medium and low dose and the preventive group have significantly lower sperm abnormality rate compared with the control group treated with lead acetate(P<0.05). There were no significant differences among high dose group, medium dose group and preventative group in sperm aberration rate (P>0.05). Sperm aberration rate of the high dose group, medium dose group and preventative group was lower than that of low dose group (P<0.05).8. Effect of recovering the anti-oxidizing systemThe GSH-Px activity of low-dose, medium-dose, high-dose group and preventative group in liver and kidney was higher than that of lead-exposed group, but lower than that of negative group(P<0.05). The GSH-Px activity of negative group and low-dose group, medium-dose group, high-dose group and preventative group in brain was significantly higher than that of model group (P<0.05), but no significant differences existed among the former four groups (P>0.05).The SOD activity of low-dose, medium-dose, high-dose group and preventative group in liver ,brain and kidney was higher than that of lead-exposed group, but lower than that of negative group(P<0.05), but no significant differences existed among the former four groups(P>0.05).The content of MDA of low-dose, medium-dose, high-dose group and preventative group in liver, brain and kidney was lower than that of lead-exposed group (P<0.05), but no significant differences existed among the former four groups and negative group (P>0.05).9. Effect of recovering the Liver function and renal functionTotal protein and albumin content in negative control group was significantly higher than that of low-dose group, medium-dose group, high-dose group and preventative group (P<0.01) and lower than that of model group (P<0.01). There were no significant differences in total protein and albumin content among all Lead-excretion Glycan-treated groups (P>0.05).The ALT, AST activity in low-dose group, medium-dose group, high-dose group was significantly lower than those in model group (P<0.05), higher than those in negative group (P<0.05). The ALT, AST activity in preventative group was lower than that of model group, but had no significant differences in comparison with negative group(P>0.05). No significant differences of ALT, AST activity existed among low-dose group, medium-dose group and high-dose group (P>0.05). There were no significant differences among all groups in ALP activity (P>0.05).The content of Blood urea nitrogen(BUN) and uric acid(UA) of high, medium, low group and preventive group was lower than those of model group significantly(P<0.05), higher than those of negative group(P<0.05). There were no significant differences among various groups except model group in the content of Blood urea nitrogen (BUN) and uric acid (UA) (P>0.05).10. Pathomorphological observation of the liver, kidney and hippocampus and malformation of spermThe liver, kidney and hippocampus damage had been found among lead-poisoned mice in the pathological examination, significant recovering could be observed in the Lead-excretion Glycan-treated groups. Lead-excretion glycan could inhibit sperm abnormalities caused by lead in mice to some extent.ConclusionsLead-excretion Glycan is a safe health food with low toxicity. It has little genetoxic and little reproductive toxicity. Lead-excretion Glycan can effectively eliminated lead without removing essential element calcium, magnesium, iron, zinc, copper, manganese; Lead-excretion Glycan can diminish the lead-induced mutagenicity and reproductive toxicity. It can also enhance the activity of the antioxidant enzyme, reduce peroxidization of lipid, remarkably recover the lead-induced liver and renal function, and inhibit pathological damages of liver, kidney and Hippocampus significantly. |
PDF Full Text Request