The Study On Absorption Enhancing Effects,safety And Mechanism Of PAMAM Dendrimers On Effective Components Of Traditional Chinese Medicine

PAMAM Dendrimer is one of the most widely studied Dendrimers in recent years,which has a large number of functional groups on the surface and cavities inside,specific three-dimensional structure and good biocompatibility,permeability and stability.It has been widely applied in drug and gene carriers fields and may become a new absorption enhancer.In order to study the absorption effect and mechanism of PAMAM Dendrimers with different concentration and different functional groups,three kinds of typical traditional Chinese medicine active ingredients-Jujuboside A,liquiritin,Evodiamine were selected as model drugs from the established Changbai Mountain traditional Chinese medicine sample library.Firstly analysised the physical and chemical properties of model drugs,secondly studied the absorption characteristics of PAMAM Dendrimers to model drugs from two aspects of in vitro and in vivo,finally explained the mechanism of PAMAM Dendrimers by DNA microarray technology,Western blot and quantitative real-time PCR technology.All of these were in order to find a wide used,high efficiency,low toxicity,clear mechanism of absorption enhancers to improve the bioavailability of drugs and change the current utilization of traditional Chinese medicine.The specific research contents are as follows: in vitro,the transport volumes of three active components of traditional Chinese medicine with or without PAMAM Dendrimers were analyzed by Caco-2 cell model,and the cytotoxicity of PAMAM Dendrimers on Caco-2 cells was analyzed by MTT experiments.The results showed that both PAMAM-NH2 and PAG(PAMAM-co-OEG)could significantly improve the transport volumes of insoluble components of traditional Chinese medicine,at the same time 0.1%(W/V)PAG had little effect on the survival rate of Caco-2 cells.In vivo,the changes of blood drug concentrations of three active components of traditional Chinese medicine containing or not containing PAG at different time points were tested after in situ intestinal loop administration,and the cytotoxicity of PAG on small intestine was analyzed by BCA and LDH experiments.The results showed that the blood drug concentrations of PAG groups were significantly higher than that of groups without PAG at each time point,and PAG could improve the absorption of insoluble components of traditional Chinese Medicine safely and significantly.Finally,the mechanism of PAG was studied by DNA microarray technology,Western blot and quantitative real-time PCR technology,and the results showed that PAG could promote the absorption of drugs by affecting clathrin in endocytic pathway to change the properties of membrane.This topic systematic studied the toxicity,promoting absorption and mechanism of PAMAM Dendrimers from the animal,cell and molecular levels,and found that PAG could safely,effectively promote the absorption of insoluble components of traditional Chinese medicine by effecting on the clathrin in endocytic pathway.Therefore,PAG is suitable to be a new type of absorption enhancer and further develops into a new type of pharmaceutical excipients.