Studies On The Reproductive And Developmental Toxicity Of Neohesperidin

Purpose:Previous studies indicated that Neohesperidin(Neo)had potential therapeutic effects in hyperglycemia and hyperlipidemia.This study systematically examined the reproductive and developmental toxicity of Neo,including adverse effect on gametogenesis,mating,pregnancy.delivery and lactation,and the adverse effects on filial generation during development.This study provides laboratory data for further investigation and application of Neo.Methods:(1)The impact of Neo in the fertility and early embryonic development of ratsVehicle control group(5%CMC-Na).Neo 0.45 g/kg group,Neo 0.9 g/kg,Neo 1.8 g/kg,and Cyclophosphamide(CP)0.02 g/kg group were adopted with 20 animals per gender in each group.Vehicle control group and 0.45,0.9,1.8 g/kg Neo group were administrated i.g.with corresponding test substances at 10 ml/kg.0.02 g/kg CP group was administrated by subcutaneous injection once a week and pregnant female rats were injected on gestation day 3(GD3)and GD6 where 2 ml/kg was used.Male rats were administrated with relevant reagents from 4 weeks before mating period to the end of mating period.Female rats were with relevant reagents administrated from 2 weeks before mating period to GD6.During the study,clinical symptoms were observed;body weight and food consumption were detected.At the end of mating period,male rats were sacrificed for gross anatomical and histopathological observation as well as sperma test.Pregnant female rats were sacrificed at GD15 for gross anatomical and histopathological observation as well as examination of uterine contents.(2)The impact of Neo in embryo and fetal development in ratsVehicle control(5%CMC-Na)group,Neo 0.45 g/kg group,Neo 0.9 g/kg,Neo 1.8 g/kg and CP 0.012 g/kg group were adopted and at least 20 pregnant female rats were used in each group.Vehicle control group and Neo 0.45,0.9.1.8 g/kg group were administrated i.p.with corresponding test substances used at 10 ml/kg by from GD6 to GD15.0.012 g/kg CP group was administrated by subcutaneous injection on GD12 where 2 ml/kg dosage was used.During the study,clinical symptoms were observed,body weight and food consumption were detected.Pregnant female rats were sacrificed at GD20 for gross anatomical and histopathological observations,and uterine contents as well as fetus examination including external,skeletal and visceral alterations were measured.(3)The impact of Neo on perinatal period in ratsVehicle control(5%CMC-Na)group,Neo 0.45 g/kg group,Neo 0.9 g/kg,Neo 1.8 g/kg and CP 0.012 g/kg group were adopted and there were at least 20 pregnant female rats in each group.Vehicle control group and 0.45,0.9.1.8 g/kg Neo group were administrated i.p.with corresponding test substances used at 10 ml/kg capacity from GD15 to postnatal day 21(PND 21).CP 0.012 g/kg group was administrated by subcutaneous injection at GD15,PND1,PND6,PND12 and PND18 where 2 ml/kg dosages were used.For maternal rats,clinical symptoms were observed;body weight and food consumption were detected and rats were sacrificed on PND21.For the first filial generation(F1),clinical symptoms were also observed;body weight and food were detected as well as developmental examination including physical development,nervous reflex,quadruped locomotion,balancing and coordinating ability,autonomic activities,learning and memory,reproductive function and histopathological examination.Results:(1)The impact of Neo on fertility and early embryonic development in ratsWhen male rats were administrated with 0.02 g/kg CP,no obvious toxicity was found on clinical observations such as body weight,food consumption,sperm motility,sperm enumeration and genital organs,but abnormal sperm incidence obviously increased.When female rats were administrated with 0.02 g/kg CP,no impact was found on corpus lutea generation before and during implantation,but significant toxicity was noted on embryonic development after implantation.When SD rats were administrated with Neo 0.45,0.9,1.8 g/kg,no obvious toxicity was found on clinical observation,body weight,food consumption and weight gain during generation,reproduction function,mating,fertility and early embryonic development.(2)The impact of Neo in embryonic and fetal development in ratsWhen pregnant rats were subcutaneously injected with 0.012 g/kg CP at GD12,body weight began to lose from GD15,the trend of which was in accordance with uterus,fetus weight and size.Meanwhile,obvious toxicity was found in external,skeletal and visceral examination.When pregnant rats were administrated with Neo from GD6 to GD15,no obvious maternal toxicity was found.Major toxicity on embryo and fetus was delayed parietal and sternebra 6 ossification.Other possible impacts might be the ossification pattern of rib,development of brain and esophagus and the ossification of mandible,cervical,lumbar,pubis,occipital and sternebra 1,3,4.(3)The impact of Neo in perinatal period in ratsWhen maternal rats were administrated with CP,the major toxicity was the decline of body weight and food consumption after delivery.For F1 generation animals,there might be some toxicity on survive,physical development,balance and coordinate ability,organ cause and function of reproductive system.When maternal rats were administrated with 0.45,0.9,1.8 mg/kg Neo,no obvious change was found on body weight,food consumption,delivery and lactation.For F1 animals,no obvious toxicity was found on survive,physical development,balancing and coordinating ability,autonomic activities,learning and memory,and reproductive function.Conclusion:Under the conditions of our current project,when SD rats were administrated with 0.45,0.9,1.8 g/kg Neo,no obvious toxicity was found on fertility,early embryonic development as well as perinatal periods,while certain extent of toxicity was found on embryonic and fetal development and the major toxicity might impact on the development of brain and esophagus along with delayed skeletal ossification.