| Objective： Investigate the protective effect of garlic polysaccharide against alcoholic braindamage on mice and research the mechanism, thus provide scientific basis to promote theapplication of garlic polysaccharides in preventing chronic alcoholic brain damages.Method： Healthy KM mice were first feed adaptively for10days, then randomly divided themice based on body weight into Control groups, Model Groups and High Garlic Polysaccharidegroups (250mg/kg), Medium Garlic Polysaccharide groups (200mg/kg) and Low Dosage GarlicPolysaccharide groups (150mg/kg), each group consists of10mice. Mice of the control groupwere perfused with distilled water. Mice from other groups were applied incremental feedingtechnique to perfuse the56degree “ErguoTou Spirit”,0.05ml/10g*Day for first3days,0.08ml/10g*Day for5days,0.10ml/10g*Day for a week and0.12ml/10g*Day till the day beforethe test ends, the dosage of0.15ml/10g*Day was used for impact gastric perfusion. Everyday30min before the spirit was perfused, distilled water was perfused for mice from control group andgarlic polysaccharide for the test groups, continue for10weeks. During the test the body weightof mice, food intake and mental status were recorded; Water maze test was proceeded in week8and week10. In week10all the mice’s body weights were measured prior to their execution, thebrains were weighed and some brain tissues were reserved in10%formalin solution forHistopathology examination. Homogenate on the brain tissues left were washed away withfreezing saline then the tissues were used for ATP enzyme, TchE, MAO, Hydrogen Peroxide,SOD, GSH-Px, CAT and MDA activity test.Result：1. General status of alcohol intoxication on miceMice of the model group showed symptoms of drool from the2ndweek, movements likescratching heads increased, bad mental status and decline in resistance also emerged. From the4thweek, mice from model group showed inflexible posture, slow in movements and terriblemental status20min after the alcohol intake, recovery happened2-3hours later. From the7thweek, mice from the model group showed chronic alcoholic intoxication symptoms like reelinggait and drowsiness, the time required for recovery elongated to6hours. When comparing thesemice to that of normal groups, they had worse appetite, low growth rate and bad fur glossiness. For the mice in Garlic polysaccharide groups, their symptoms of intoxication were better and thetime required for recovering is shorter, about2-3hours, also they had better appetite and furglossiness than that of mice from model group. These results demonstrated that garlicpolysaccharides are helpful in mend the adverse effects of alcohol intoxication.2. Comparison of body weight and brain weight change of mice from different groups.From week1to week3the body weight of mice from all groups show an obvious tendency ofincreasing, from week4to the end of the whole experiment the body weight of mice from modelgroups were decreasing slowly all the time. The final average body weight is statisticallydifferent from that of normal groups (P<0.05), which means that long term excessive drinkingwill affect the appetite of mice and cause malnutrition on the mice. Compared to model groups,body weight of mice from garlic polysaccharide groups retained relatively stable from week5and possessed a slow increasing tendency, from week5the body weight of mice from lowdosage garlic polysaccharides group showed statistical difference from that of model group(P<0.05), as for the body weight of medium and high dosage garlic polysaccharide groups, thisphenomenon happened in week6and the body weight change of medium dosage group wasmore volatile.Combined with the change of mice’s appetite from all the groups, the test figured out that whencomparing to normal groups, mice’s appetite from model group had been declining from week3to the end of the whole test, as for the mice’s appetite from the garlic saccharide groups, thedecline weren’t that obvious. The result prompted that garlic polysaccharide could ameliorate theappetite of mice.Compared to the normal group, the brain weight index of model group is higher but thedifference has no statistical significance, this result prompted that the brain damage from alcoholon mice still remains preliminary and no organic lesion happened, the mechanism still require afurther research to find out. Compared to model groups, the brain weight index from garlicpolysaccharide groups were lower and very close to that of normal group.3. Morris water maze test analysis.Morris water maze could determine the mice’s ability to obtain and retain memory viadetermining the escape latent period and searching performance. We’ve tested the mice in week6thand week10th. The result in week6revealed that, after4days of fixed location swimming test, the timerequired for mice from all test groups to successfully escape had shortened. Compared to normalgroups, the mice from model groups had statistically longer escaping latent period from day2.Compared to model group, mice from all garlic polysaccharide groups had statistically shorterescaping latent period (P<0.05), but their space searching ability remain almost undifferentiated(P>0.05), these phenomenon indicated that six weeks’ continuous alcohol intake had alreadydone damage on the mice’s memory obtaining ability, but their memory retaining abilityremained unaffected, such alcoholic effects on mice’s brain might be still preliminary and has thepossibility for recovery, garlic polysaccharide has certain protective effects on brain againstalcoholic damage.In week10, the mice’s fixed location swimming test resulted same escaping latent period to thatof week6, confirmed that the memory obtaining ability of mice from model group was damaged.Except for mice from high dosage garlic polysaccharide group, mice from other dosage groupspossessed statistically shorter escaping latent period compared to week6(P<0.05), also onceagain confirmed the protective effects of garlic polysaccharide on brain.Meanwhile, the space searching ability of mice from all groups began to vary obviously, normalgroups was better than model group and high dosage garlic polysaccharide group was better thanmedium, low dosage and model group. This confirmed that garlic polysaccharide has protectiveeffects on mice’s brain against alcoholic memory damage, but no dosage-effect relation wasshowed, the best dosage and effective threshold still require more research to investigate.4. Neurotransmitter enzyme index comparison of all groupsCompared to normal groups, the Na+-K+-ATP activity of model group declined significantly, butthe activities of TchE and MAO increased significantly (P<0.05), which demonstrated thatalcohol has the ability to prohibit the activity of MAO and TchE. Compared to model group,garlic polysaccharide groups had obvious higher Na+-K+-ATP activity, the MAO and TchEactivities of low and medium dosage groups declined, these results means that garlicpolysaccharide has the ability to regulate the unusual rise of activity of MAO, TchE and theNa+-K+-ATP activity decline caused by alcohol, but the effective dosages are different.5. Comparison of various brain damage indexes from different test groups.Compared to normal groups, the SOD, GSH-Px activity of model group declined(P<0.05), MDA content increased(P<0.05); Compared to model groups, SOD, GSH-Px of different garlicpolysaccharide groups increased obviously(P<0.05), MDA content declined(P<0.05), thehydroxyl peroxide content in brains of these groups were almost the same. For the CAT activity,model groups had lower activity than normal group（P>0.05）, garlic polysaccharide groups hadhigher activity than model group（P>0.05）。For the H2O2activity, model groups had loweractivity than normal group（P>0.05）, garlic polysaccharide groups had higher activity than modelgroup（P>0.05）。6. Pathological tissue change comparison of mice from all groups.From the hippocampal convolution section we can see that compared to normal groups, the micefrom model groups had showed more cells’ necrosis and edema, but the mice from three garlicpolysaccharide groups were almost identical to normal group, showed few cell necrosis, whichshowed that garlic polysaccharide could notably decrease the neuron damage and protect thebrain tissue.Conclusion：1, the increasing doses of the stomach for alcohol, filling method can be successful preparationchronic alcoholism small brains damage model, this model can be used for chronic alcoholismbrain damage mechanism research, as well as to the prevention and control of drug and healthfood role of functional factors, development and selection;2, and sustained long-term excessive drinking can lead to small brains damage and learning andmemory neural behavioral change;3, garlic polysaccharide on mice in brain tissue damage chronic alcohol toxicity have obviousprotection;4, garlic polysaccharides have reduce chronic alcoholism mice brain nerve neurotransmitterTchE and MAO activity effect, so as to improve the choline can and single amine the function ofnervous system.5, garlic polysaccharide can by improving the chronic alcoholism mice brain tissue antioxidantenzymes SOD, GSH-Px activity, scavenging free radicals, restrain lipid peroxidation, reduce thecontent of MDA, thus preventing alcohol on mouse liver tissue damage effect. |
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