Study On Safety Evaluation Of Pearsonothuria Graeffei Saponins

Saponins are the main secondary metabolites of sea cucumber, and have various physiological and pharmacological activities of anti-tumor, anti-bacterial, improving immunity, but also has a strong cytotoxic effect of hemolysis, liver and kidney toxicity. At present, more than100kinds of saponins monomers are separated from sea cucumber. In this study, taking Pearsonothuria graeffei(P. graeffei) as the research object because of its high content of total saponins, first of all acute toxicity test is used to confirm median lethal dose(LD50) and toxicity grade. On the base of this, a series of tests are conducted refered to the toxicological safety evaluation procedures and methods including Ames test, mouse bone marrow cells micronucleus test, mouse sperm malformation test, thirty-day feeding test, teratology testing and ninety-day feeding test. The main results for this study are as follows:In acute toxicity test, different doses of P. graeffei saponins were given to rats by one time, and conditions of poisoning and death in rats were observed within14days after administration. The calculated LD50was0.797g/kg and0.818g/kg in male and female respectively and it belonged to a low toxicity level according to toxicity criteria. P. graeffei saponins had no remarkable difference on the blood cell content and serum biochemical indexes compared with the control group, which indicated that there was no significant damage to kidney and liver in rats.Genitic toxicity studies were conducted on the basis of LD50, including Ames test, mouse bone marrow micronucleus tests and sperm deformity test. Ames test results were negative indicating that P. graeffei saponins had no mutagenic activity to any of the test strain. Furthermore, based on the observation of bone marrow smear in bone marrow cells micronucleus test, the result showed that no significant difference in micronucleus rate, which indicated that135-540mg/kg (1/8-1/2LD50) saponins have no genotoxic effects to marrow cells. In sperm teratogenicity test in mice, the result showed that significant deformity changes were observed only at540mg/kg in the epididymis by observing the sperm deformity rate, which indicates that540mg/kg (1/2LD50) saponins act as genotoxic effects on the male reproductive cells.For teratogenicity study, pregnancy rats were treated with different doses of P. graeffei saponins, the number of total fetuses, living fetuses, dead fetuses and absorbed fetuses were recorded. The weight of uterus, placenta,fetuses and length of body, tail were measured and appearance, internal organs and skeleton of fetal rats were tested. The result revealed that there was no difference between the control and treated groups in terms of reproductive parameters.This suggested that P. graeffei saponins had no adverse effect on development, appearance, skeleton and viscera of the fetuses which gave evidence to no embryonic toxicity and teratogenicity of12.3-192.5mg/kg saponins.For subacute and subchronic toxicity test, different doses of P. graeffei saponins doses were given to the rats of treatment groups for30and90days, test indices including body weight, blood cell content, blood biochemical index and histological observation. The results indicated that no obvious changes in each blood index were observed compared with the normal group, but obvious toxicity damage was found in testis in the high dosage group.100mg/kg P. graeffei saponins induced subacute toxicity to the main target organ of testis.30d-feeding test findings indicated that significant morphogenesis changes were observed of rats testis, sperm cells levels reduced and degradation degenerated in high dose of100mg/kg(12.8%LD50)saponins which preliminarily demonstrates P. graeffei saponins have certain subacute toxicity in male rats.90-day toxicity study shows further evidence that100mg/kg saponins have subchronic effects in male rats, and testis is the main toxic target organ.In conclusion, there are no toxic effect in rats under the dose of50mg/kg-bw and the safe dosage is less than8mg/kg-bw converting to human dose. The results offer a theoretical basis and scientific basis for the development and utilization of sea cucumber saponins and promote the development of deep-processing industry of sea cucumber.