| OBJECTSThis research through the toxicological experiment and auxiliary cholesterol-lowering experiments to study the effect of burdock tea in reducing blood lipid and its safety, and provide a scientific evidence for the utilization of burdock in functional food, health care products, drug and other related areas.METHODS1Toxicology Experiment(1)Acute toxicity test Male and female each half, weighting18.2-21.6g,20mice,each mice was given0.2ml/10g.bw.Every day mice were gavage fed twice and the infected dose was20.0g/Kg·bw. Continous observation for14days,then the mortality and performance were recorded.(2)Hereditary toxicity test①The mutation of salmonella test (Ames test) In accordance with the experimental results of the toxicity,eight groups were set up. There were five doses (8,40,200,1000,5000μ.g/dish) groups and the other three groups were the spontaneous back to become group, solvent control group and positive control group.Experiments were done with and without S-9mixture on the basis of the flat mixing method.The experimental strains of TA97,TA98, TA100,TA102are histidine defected rat typhoid salmonella that fulfilled the requirements. The vitro activating systems was S-9mixture which was rat liver homogenate that caused by polychlorinated biphenyls(PCBs).In each group,three dishes were set up. It was judged to be positive as follows.If back to become number of bacterial colonies about test sample is twice as much as the spontaneous number, meanwhile there was a relationship of dose-response. In the same conditions, the methods were repeated twice.②Micronucleus Test of Mice Bone Marrow Polychromatic Erythrocytes Male and female each half, weighting25～30g,50mice were divided into5groups randomly, each group had ten mice. The negative control group were given distilled water, and the positive control group were given cyclophosphamide (40mg/Kg·bw).These groups were given different infected doses of2.5,5.0,10.0g/Kg·bw. The feeding amount was0.2ml/10g.bw and mice were gavage fed twice with an interval of24h.After6hs the second fed, mice were killed,the femoral production was made and then did microscopic examination.Under the microscope, every mice was detected1000polychromatic erythrocytes. Micronucleus cells’number was recorded.And the rate of micronucleus was calculated (micronucleus cells/PCE cell, by thousands of points).200PCE were detected, normochromatic erythrocytes’number and ratio was calculated.③Mouse sperm deformity experiment Weighting25-30g,25male mice,were randomly divided into5groups, each group had five mice, grouping and processing like②Three groups were given different infected doses of2.5,5.0,10.0g/Kg·bw. The negative control group were given distilled water. Besides the positive control group were given40mg/Kg·bw cyclophosphamide. Every day mice were gavage fed one time. Gastric irrigated constant for5d once a day. After35d the first gastric irrigation, mice were killed.Besides both sides of the epididymal production was made and did microscopic examination.Each mice was found1,000sperms by structural integrity, the number and type of distortion were recorded, the rate of sperm abnormality were calculated (by percentage).(3)30d feeding test in rats Male and female each half, weighting74～94g,80weaning rats,were randomly divided into four groups. Each group had twenty mice. According to the human body recommended set up three groups, given different doses of3.75.7.50.15.00g/Kg·bw.(equivalent to the recommended dose of25.50,100 times). The negative control group, gave the same volume of distilled water. Single cage feeding,free food,continuous observation30d.drinking water but fasting continued for16h, and then biochemical tests was detected. In the meanwhile the pathological examination on visceral was also checked up.2Auxiliary cholesterol-lowering experiments40weaning rats, male, weighting74～94g. According to the blood lipid lever, mice were randomly divided into four groups, each group had ten mice. The groups were divided into high fat control group, low dose group, middle dose group, high dose group respectively. High fat diet and gavage fed distilled. Low, middle, high dose groups were given high fat diet and gavage fed different dose of test substance.30d later, took stern blood and tested blood lipid index.RESULTS1Toxicology Experiments(1)Acute toxicity test During the test,mice have no symptoms of poisoning performance and death. According to the MTD results, the biooxyrate content on the female and male mice were more than20.0g/Kg·bw. On the basis of the acute toxicity grading standard, the sample of acute toxicity is non-toxic level.(2)Hereditary toxicity test①The mutation of salmonella test (Ames test) In each dose group, the back to become number of bacterial colonies was no more than the spontaneous number, and no dose-response relationship was observed either in two experiments. The result of the test is negative. It is indicated that with and without S-9have no genetic toxicity effect on rat typhoid salmonella TA97, TA98, TA100, TA102.②Micronucleus Test of Mice Bone Marrow Polychromatic Erythrocytes In comparison of the negative control group, the micronucleus rate in each experimental group has no significantly different (P>0.05), but positive control group was significant difference (P<0.01). It is illutrated that this sample can not cause the micronucleus mutation of rat’s bone marrow polychromatic erythrocytes.③Mouse sperm deformity experiment In comparison with the negative control group, in each experimental group mice sperm deformity rate has no significantly different (P>0.05).But positive control group has significant difference (P<0.01).It is illustrated that the test substances can not cause sperm abnormality for mice.(3)30d feeding test in ratsThe experimental results show that each experimental group animals’growth is good in the test cycle. Compared with the control group, there were no significant difference (P>0.05) of all the index of weigh gain, food utilization, blood routine and blood biochemistry index, viscera weight and viscera coefficient. And the index was within the normal range in this experiment. This illustrates that each experimental group without poisoning symptoms and death.2Auxiliary cholesterol-lowering experiments1n the whole experimental period. The level of total cholesterol (TC) and triglyceride (TG) of the low、middle、high dose groups is lower than the control group((P<0.01or P>0.05)The level of high denity lipoprotein cholesterol (HDL-C) is higher than the control group(P<0.05)The level of TC goes down by0.32%,15.3%,21.4%in different dose groups. The level of TG decreases by5.9%,22.6%,32.4%in different dose groups. The level of HDL-C raises0.02mol/L、0.24mmol/L、0.32mmol/L in different dose groups.CONCLUSION1. In acute toxicity,the mice had no symptoms of poisoning performance and death. According to the MTD results, the biooxyrate content on the male and female mice were more than20.0g/Kg·bw. On the basis of the acute toxicity grading standard, the burdock tea is non-toxic substance.2.By the hereditary toxicity test including the Ames test,micronucleus test and sperm deformity experiment results were negative.lt is indicated that under the test conditions burdock tea have no hereditary toxicity.3. The result of the30d feeding test in rats. Burdock tea for the observation indexes of rats did not have obvious toxic effect, in the recommended range using the burdock tea is safe. 4. According to ’technical specifications for health food inspection and evaluation’, Burdock tea have auxiliary cholesterol-lowering function. Because that different doses of Burdock tea can increase HDL-C level reduce TC and TG levels. |
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