| Objective:this study aims to observe the influence of different doses of puerarin to the genital system of sexually matured female Kunming mice and the embryo-fetus development of the mice, and explore the mode of puerarin to the mice in reproductive period and its genotoxicity, providing experimental basis for the clinical application of puerarin.Method:1) observed the estrous cycle through the vaginal cytology smear in mice and change of the uterus through uterotrophic assay, surveyed the mouse's estradiol follicle-stimulating hormone,and luteinizing hormone and so on through enzyme-linked immunosorbent assay, to evaluate the influence of puerarin to the genital system of sexually matured female Kunming mice. Chose 40 female Kunming mouse of 8-9 weeks. divided the chosen rats into 4 groups according to the random digits table:low dose puerarin group (150mg/kg),high dose puerarin group (450mg/kg),diethylstilbestrol group (35mg/kg) and solvent control group(sodium carboxymethyl cellulose), all groups were given medicine by intragastric administration one time a day with 0.2 ml every time. Observed the vaginal cytology smear everyday at 8:00. Weigh the mice weekly weighing and adjusted the dose of drug usage according to the weight. After 6 weeks of continuous administration and during the 24 hours before the rats' diestrums after the last administration, took the Orbital blood and kill the mice off the cervical spine. Took the uterus and ovary's wet weight quickly to calculate. All serum samples were stored at -20℃, one side of the ovary, uterus were quickly moved to -80℃degree for preservation, the other side of the ovary and uterus were fixed in 10% formalin, and made paraffin production of HE staining for observing uterine pathological changes.2) Treated the pregnant mice with different doses of puerarin since the implantation of the embryo to the hard palate closure period (6-15 days of pregnancy mice), and assessed the influence of puerarin on female pregnant mice, embryonic and fetal development of the fetus. The experiment used 160 8-9 week old mice, male and female both for half, divided the female rats into 4 groups according to the random digits table with 20 rats one group. The mice of the same month and variety were caged together at 1:1 ratio on sex. and from the next day. examined whether there were vaginal plug in the female mice's vagina, and examined the mice without vaginal plug whether there were viable sperms. If vaginal plug or sperms were found. they were designated as pregnancy"Od",then the mice were fed for 2 weeks and given intragastric administration from the sixth to fifteenth day in pregnancy. Killed the mice off the cervical spines at the eighteenth day. Separated the fetal rats from the top of the uterus, checked visually for dead fetal and absorbed fetal, calculated the number of live births and checked if there were deformities in their shape,skeleton or abodominal organs. Calculated the average weight of live fetuses, measured the fetus'average top-rump length with a vernier caliper.Result:1) weight:the low dose puerarin group were significantly lower than the normal control group (P<0.05). also significantly lower than the high dose puerarin group and the diethylstilbestrol group (P<0.01). there were no obviously significance among the high dose puerarin group,the diethylstilbestrol group and the control group.2) Organ coefficient:ovary coefficient, there were no obviously significance among the four groups; uterus coefficient. the diethylstilbestrol group were significantly higher than the low dose puerarin group and the normal control group (p<0.01). and significantly higher than the high dose puerarin group (p<0.05), the low dose puerarin group is lower than the control group, the high dose puerarin group is higher than the control group but no significant difference.3) The rats'oestrous cycles:we computed the rates of cycle disordering of the 42th day, the disordering rate of the four groups were respectively 70%,80%,100%,0, then it can clearly be seen that low dose puerarin,high dose puerarin and diethylstilbestrol all can lead to the rats'oestrous cycles disordering, the low dose puerarin group and the high dose puerarin group were significantly lower than the diethylstilbestrol group.4) Serum E2:the low dose puerarin group were significantly lower than the normal control group (P<0.05).and also significantly lower than the high dose puerarin group and the diethylstilbestrol group (P<0.01). the high dose puerarin group and the diethylstilbestrol group were significantly higher than the normal control group (P<0.01 the high dose puerarin group were significantly lower than the diethylstilbestrol group (P<0.01).5) Serum PSH:the low dose puerarin group were significantly lower than the other three groups (p<0.01), the high dose puerarin group and the diethylstilbestrol group were significantly higher than the other two groups (p<0.01), there was no significant difference between the high dose puerarin group and the diethylstilbestrol group.6) Serum LH:the low dose puerarin group were significantly lower than the other three groups, the high dose puerarin group and the diethylstilbestrol group were significantly higher than the other two groups (p<0.01), there was no significant difference between the high dose puerarin group and the diethylstilbestrol group.7) Histopathological examination of the uterine:the uterines of the diethylstilbestrol group were fat, and the interstitial substance of the endometrium were dropsical, the glands increased, and were flexural with luminal expansion and different forms. Glandular epithelial cells were tall columnar pseudostratified with slight thickening of the basal layer. The other three groups didn't have these changes.8) Endometrial thickness:the diethylstilbestrol group were significantly higher than the other three groups (p<0.01), and there was no significant difference between the other three groups.9) the pregnant mice's weight:there was no significant difference among the weight of the 6th,9th,12th,15th day, the diethylstilbestrol group were significantly lower than the other three groups, and there was no significant difference between the other three groups.10) Formation of mouse embryos:the number of live fetus, the low dose puerarin group and the high dose puerarin group were lower than the other normal group, but no significant difference, and were significantly higher than the diethylstilbestrol group (p<0.05), there was no significant difference between the high dose puerarin group and the low dose puerarin group. The diethylstilbestrol group were significantly lower than the control group (p<0.01); there was no significant difference in the number of absorbed fetus; there were no dead fetus in the four groups.11) Situation of fetal development:the average weight, the low dose puerarin group were lower than the high dose puerarin group and the control group but with no significant difference, the high dose puerarin group were higher than the diethylstilbestrol group and lower than the control group but with no significant difference. the diethylstilbestrol group were significantly lower than the control group (p<0.01):the average top-rump length, the low dose puerarin group were significantly lower than the control group and the high dose puerarin group. and higher than the diethylstilbestrol group but with no significant difference. the high dose puerarin group were higher than the diethylstilbestrol group and lower than the control group but with no significant difference, the diethylstilbestrol group were lower than the control group (p<0.01)Conclusion:1) Under the experimental conditions, puerarin have certain influence on the genital system of the sexually matured mice, different doses of puerarin play dual role of antiestrogen and antiandrogen.2) the pregnant female rats after administration had no significant adverse effects to the pregnant rats and their offspring, that is to say, puerarin has no clear embryo-fetus developmental toxicity.
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