The Study On Anti-damage Of Inonotus Polysaccharides On The Pregnant Mice Placental Infected By Bovine Neospora Caninum

Neospora caninum(N.caninum)is a kind of protozoan which inhabits in the cells of dog,cattle,sheep and other mammalian animals.N.caninum disease can be vertically transmitted via the placenta and seriously injury the placenta of pregnant domestic animal,cause female abortion,stillbirth or neonatal dyskinesia and nervous system diseases.N.caninum disease seriously harms to the cattle and has become one of the main factors endangering the healthy development of cattle industry.At the present time there are no specific drugs and vaccines for the prevention and control of N.caninum.In this study in vitro in vivo insect resistant test,pathology observation,placental apoptosis gene and some indexes affecting material transport of pregnant rats placenta,placental blood flow and the growth and development of the fetal were detected in order to explore the protective effects of Inonotus obliquus polysaccharide on the placenta of pregnant rats infected by N.caninum.This study will provide theoretical and experimental basis for development and utilization of drugs resisting the N.caninum.1.The experiment of Inonotus obliquus polysaccharides resisting the tachyzoite of N.caninum in vitroIn this experiment,the infected cell model in vitro was established by the normal Vero cells infected by N.caninum from cattle.The toxicity test and insect resistant test Inonotus obliquus polysaccharides cell was conducted by the method of MTT.By comparing the cell proliferation rate,the insect-resistant effect and the best dosage in vitro of the drug was determined.The results showed that when the concentration of Inonotus obliquus polysaccharide is 100 ug/mL.the cell proliferation rate was the highest and was 37%and 48%,respectively at 24 h and 48 h,and the insect-resistant effect was the best and had no cell toxicity.2.The observation of the protective effects of Inonotus obliquus polysaccharide on pregnant mice placenta infected by N.caninumIn this experiment,pregnant mice models infected by N.caninum from cattle were used to determine of the effect of Inonotus obliquus polysaccharides at different concentrations on live births,placental weight and placental factor and the damage of the pregnant mice placenta tissue.The results showed that polysaccharides in the high and medium dose group could reduce the living fetus number of pregnant mice,obviously increase the placental weight and placental ratio and reduce the damage of the structure of placenta caused by N.caninum from cattle.The difference was very significant compared with model group(P<0.01).3.The effect of Inonotus obliquus polysaccharide on apoptosis gene expression of pregnant mice placenta infected by N.caninumIn this experiment,the total RNA of the mouse placenta was extracted,and the PCR chip was used to detect the placental apoptosis genes and these genes were compared.The results showed that The results showed that 66 genes were detected by qRT-PCR,and between Inonotus obliquus polysaccharide and the infected group,63 genes were up-regulated and there are 8 differentially expressed genes in them,namely BCL2.TNFRSF11B,RIPK1,BIRC2(C-IAP2),BCL2A1,BCL2A1,DAD1 and BNIP2,and 3 genes were down regulated.4.The effect of Inonotus obliquus polysaccharide on material transport of pregnant mice placenta infected by N.caninumIn this experiment,CG,PL,E3 and ACH hormones in the serum of pregnant mice infected by N.caninum were detected by ELISA in order to determine the effect of Inonotus obliquus polysaccharide on material transport of pregnant mice placenta.The results showed that polysaccharides could up or down regulate CG,PL,E3 and ACH production of pregnant mice in high and medium dose group.The difference was very significant compared with model group(P<0.01).5?The effect of Inonotus obliquus polysaccharide on the cytokines of bloodflow of pregnant mice placenta infected by N.caninum and fetal growth anddevelopmentIn this experiment,pregnant mice models infected by N.caninum from cattle were used to determine the levels of CRH,TGF-? and IGF-2 hormones in pregnant mice placental cell suspension related to placental blood flow and fetal growth and development.The results showed that Inonotus obliquus polysaccharides in the high and medium dose group could significantly relieve CRH,TGF-? and IGF-2 secretion disorder caused by N.caninum infection.Conclusion:1.Inonotus obliquus polysaccharides played a good role on N.canimum resistance in vitro and the best effect was 100 ug/ml and no cytotoxicity.2.Inonotus obliquus polysaccharides could relieve the damage of pregnant mice placenta tissue caused by N.caninum infection and reduce miscarriage and stillbirth caused by N.caninum infection.3.Inonotus obliquus polysaccharides could change some apoptosis genes expression in pregnant mice placental infected by V.caninum.reduce the apoptosis of placental tissue cells,improve the placental trophoblastic cells atrophy caused by cell apoptosis and decrease the occurrence of abortion and stillbirth.4.Inonotus obliquus polysaccharides could improve material transfer obstacles,the blood perfusion inadequacy and fetal developmental delays caused by N.caninunm infection,ensure adequate nutrition supply for fetus and reduce the occurrence of adverse pregnancy such as fetal developmental delays,abortion and stillbirth caused by placental honnone adverse effect.