Different Effects Of PINELLIA RHIZOMA And Its Formula After Compatibled With Ginger And Ginseng On Mouse Embryonic Development And MESC Differentiation

Prenatal drug safety plays a vital role on ensuring children’s healthy gro,th and development.Traditional Chinese medicine(TCM)have long been aware that the important significance of prenatal care,and formed the practical system and theory of prenatal care,method of "pregnancy contraindication medicine" establishment being one of them.Pinelliae,a traditional "pregnancy contraindication medicine",in the use of fetal period has always been controversial,but Dried Ginger-Ginseng-Pinellia Pill,creating by medical sage,zhang zhongjing,as a treatment for pregnancy vomiting has always been in use until today.Dried Ginger-Ginseng-Pinellia Pill in the treatment of pregnancy vomiting,whether causing fetal hinder by the use of raw pinelliae?whether raw pinelliae has embryonic toxicity?With the compatibility of dried ginger-ginseng-pinellia decoction,it can be toxic attenuated or not?Solving those questions needs more objective scientific basis.This experimental research is a part of the National Natural Science Foundation of China Youth Fund Projects-"Based on proteomics methods to explore Banxia on embryonic development and compatibility impact attenuation mechanism".Using reproductive toxicity tests in sensitive period to teratogenic agent,explore the effects of Pinelliae(BXd)and Dried Ginger-Ginseng-Pinellia Decoction(GJRSBXd)on pregnancy maternal and embryonicdevelopment of mice.At the same time,through the establishment of mouse embryonic stem cells(mESC)test in vitro,using the raw pinellia decoction(BXd),dried ginger-ginseng-pinellia decoction(GJRSBXd),raw pinellia powder(BXp),dried ginger-ginseng-pinellia powder(GJRSBXp)different drug-containing serum intervention training,study the influence on mESC differentiation,in order to explore the raw pinellia effect on embryonic development and dried ginger-ginseng-pinellia formula security in the prenatal period.Objective1.To concluse the application of the embryonic toxicity study research of traditional Chinese medicine(TCM)in vivo and in vitro experiments.To conclusion study of raw pinellia,dried ginger-ginseng-pinellia compound in the progress of the embryo toxicity.2.To explore the effects of BXd and GJRSBXd on pregnancy maternal and embryonic development of mice.3.To compare differentces on BXd,GJRSBXd,BXp,GJRSBXp medicated serum cultivating mESCs differentiating into cardiomyocytes.4.To discusse the influence of the safety to the fetus when applicating raw pinellia and dried ginger-ginseng-pinellia formula treating pregnancy vomiting.Methods1.Reproductive toxicity tests in sensitive period to teratogenic agent were applied to detect the impact on gestation and embryonic development.,using SPSS 19.0 software to analyze the results.2.Classic hanging drop three-step method used to culture mouse embryonic stem cells(mESC),formating the embryoid bodies(EBs),with the different drug-containing serum of BXd,GJRSBXd,BXp,GJRSBXp,the blank group and FBS cultivating 14 day,comparing the differentiation express of mESC totipotency protein Cont3 and myocardial specificity protein TnT,using Photoshop CS5 and SPSS 19.0 software to analyze the results.Results1.The results showed that there was no obvious toxicity in mice administrated with BXd(2.275g·kg-1)or GJRSBXd(5.688 g·kg-1)for 14 days than mice in normal group in terms of weight and main organ pathology.No distinct toxicity of both BXd and GJRSBXd was found on pregnant mice and their fetus after administration for 10 days in susceptible period.2.With BXd,GJRSBXd,BXp,GJRSBXp medicated serum,blank serum,FBS cultivating EBs for 14 d,comparison Cont3 protein expression level.Compared with the FBS group,BXd group and BXp group upper expression(P<0.01),GJRSBXd group and GJRSBXp group have no significant difference;Compared with BXd group,expression of GJRSBXd group is lower,but no statistical difference;Compared with BXp group,GJRSBXp group expression is significant lower(P<0.05).3.With BXd,GJRSBXd,BXp,GJRSBXp medicated serum,blank serum,FBS cultivation EBs for 14 d,comparison TnT protein expression level.Compared with the FBS group and the blank group,BXd group has a significant lower expression(P<0.05),BXp group also has a significant lower expression(P<0.01),GJRSBXd group and GJRSBXp group have no significant difference;Compared with BXd group,GJRSBXd group has a significant upper expression(P<0.05);Compared with BXp group,GJRSBXp group has a significant upper TnT expression(P<0.01).Conclusion1.BXd and GJRSBXd do not have embryotoxicity in the dosage of this experiment,but BXd can cause pregnant rat maternal weight reduction.2.Raw pinellia drug-containing serum maybe generate embryonic toxicity by inhibiting mESCs differentiate into cardiomyocytes in vitro,while dried ginger-ginseng-pinellia formula under our experiment condition has no obvious embryonic toxicity.3.Raw pinellia as "pregnancy contraindication medicine" is not recommended using of the prenatal period alone,dried ginger-ginseng-pinellia formula with not obvious embryotoxicity,can be safely used by syndrome differentiation in fetal period.