Preliminary Study Of Development Toxicity,Target-organs Of Toxic TCM On Zebrafis

Traditional Chinese Medicines are a great treasure-house and receiving worldwide publicity for their unique efficacy.Toxic traditional Chinese medicines are an important part of traditional chinese medicines in the clinical treatment,it is of important significance in the prevention and control of diseases,therefore,making use of the toxic TCM draws the more and more researchers' attention,for example,aconitecan be used to treat the Rheumatic arthralgia by use its Paralysis efficacy.purified cobra venom to sequel of Poliomyelitis and so on.Recently,reports of adverse reactions about TCM because of their irrational use are continues to rise year over year,which have a significant effect on their reputation and international status,therefore,for the purpose of improvement and the development of the theory of traditional Chinese medicine,modern toxicologists always try to perform a security assessment for the TCM widely used in the clinic with the modern pharmacology method,furthermore,provide a series of effective and supporting data for the theory and practice of like cures like of TCM and guide reasonable clinical prescriptions.Zebrafish,in recent years,has become an alternative of animal model to study the pharmacology and toxicity of TCM.There is strong rationale to conduct the toxicity studies in zebrafish including the following its small size,low cost,large numbers of offspring,rapid development of most organs,capacity to absorb low molecular weight compounds dissolved in the culture water make it suit for small molecule screening,especially for high-throughput screening.The virtually transparent embryos of this species especially in the process of early embryonic development result in the ability to create or increase in vivo imaging capabilities.This way allows us to non-invasive observe the various stages of the growth and development of organ of the zebra-fish through an optical microscope permiting the rapid and ease evaluation of the responses of live embryos to drugs in real time and obtaining the experimental data from the multiple perspectives facilitating the evaluation of the end of toxicological experiments.as a vertebrate animal,zebrafish have a high degree of genetic conservation and their morphological and molecular basis of tissueand organ development is either identical or similar to other verte-brates including humans.The nervous,cardiovascular and visual system are highly conserved at the level of anatomical,physiology as well as molecular level.Because embryos receive nourishment from an attached yolk sac,no feeding is required for7 days post fertilization(dpf).furthermore,With,these characteristic of zebrafish discussed above combined with the mature of zebrafish anatomy,histological section technology and the omics technology has embraced zebra-fish embryos as a convenient,robust alternative in vivo model system appropriate for studies of toxicity of TCM which can greatly accelerate the drugdiscovery process,decrease costs,and provide more accurate resultsthan cell-based assays.Methods:1.Establishement ofclosed recirculating aquacultural system for zebrafish.2.Assess the effect of toxic TCM on development of zebrafish.3.Target organs of different toxic TCM on zebrafish were initially identified by The results of phenotype observation and acridine orange staining method.4.We compared results of toxic TCM in zebrafish with results for mammalian models.Result:1.Process of culture of zebrafish were practiced,the Recirculating Aquaculture System was up to relevant standards with the sufficient stability and reproducibility:fertility rate of embryos was greater than 70%,hatching rate was was greater than 90%,mortality rate was lesser than 10%.2.Different toxic TCM have different toxic effects on zebrafish development process,causing the loss or the decress of voluntary movement,heart rate and so on.3.Different toxic TCM can affect the organ of zebrafish to a variable extent.Toxic target organs of TCM was preliminarily determined with the acridine orange staining method since it can permeate through the cell membranes and label the nucleotides,cause bright green fluorescence observed under fluorescent microscope.Conclusion:In this study,we assessed the effect of toxic TCM in the chinese pharmacopoeia on the developmental toxicity,acute toxicity and confirm the correspondent target organ with the zebrafish model cultured by in our laboratory.The results show that experments had good repetition and the zebrafish are suitable for the High-throughput toxicity studies ofTCM.This study just was the preliminary study about the toxic TCM to provide a basis for the toxicity theory and the safe safe use in clinical in the future.