The Transport Study Of Four Water-soluble Derivatives Of Andrographolide On Caco-2 And Calu-3 Monolayer Models

The absorption,distribution,metabolism and excretion rule of Traditional Chinese Medicine and its active components in organism is one of the hotspots in the modernization of Traditional Chinese Medicine.It is of great significance to understand the scientific connotation of Chinese Medicine and optimize the dosage form of drugs.The current research on the absorption of Traditional Chinese Medicine and its active components is mainly based on animal experiments.The main drawback is that there are species differences,and the operation is complex,the cycle is long and the reproducibility is poor.Compared with animal experiments,the cell model in vitro effectively avoids the difference between the species of the experimental animals and the human body,and has high reproducibility.It can be used for high throughput screening and has a broad prospect for development.In recent years,more and more researchers used in vitro single cell model to evaluate the in vivo absorption of Traditional Chinese Medicine and its active ingredients.Andrographolide is the main active ingredient of Andrographis Paniculata which can be used either as a food or as a drug.Andrographolide and its water-soluble derivatives have many biological activities,such as anti-inflammatory and antiviral,and are widely used in the treatment of respiratory diseases.In this paper,the Caco-2 cell model and Calu-3 cell model were established to observe the transport of four Andrographolide water-soluble derivatives on the single cell layer model,and to predict the permeability of the drugs in the digestive tract mucosa and the respiratory mucosa.Four kinds of Andrographolide water-soluble derivatives were determined by HPLC.The results showed that these methods are suitable and accurate,and can be used for the precise determination of Andrographolide water-soluble derivatives.The MTT method was used to calculate the survival rate of Caco-2 cell line and Calu-3 cell line after 24 hours action of water soluble derivatives of Andrographolide with different concentrations.The effects on the activity of two cell lines were compared,and the safety concentration of the drug delivery was determined.The results showed that four kinds of water soluble derivatives of Andrographolide showed inhibitory effects on cell activity of two cell lines,and showed a concentration dependent manner.Four kinds of water soluble derivatives of Andrographolide inhibited cell activity of Caco-2 cell line more than Calu-3 cell line.Within a given concentration range,the safety of four kinds of Andrographolide water-soluble derivatives from large to small are Andrographolide total Sulphonates,Andrographolide Sodium Bisulfite,Potassium Sodium Dehydroandrographolide Succinate and Potassium Dehydroandrographolide Succinate.When the concentration is below 500?g·mL-1,four kinds of Andrographolide derivatives act on two cell lines,and their survival rates are close or even higher than 90%.Therefore,0-500 ?g·mL-1 is a safe drug concentration during the transshipment experiment.The Caco-2 and Calu-3 single cell models were successfully established,and the effects of concentration,time and P-glycoprotein inhibitors on the transport of Andrographolide water-soluble derivatives were investigated.The results showed that the bilateral transport of four Andrographolide water-soluble derivatives on the Caco-2 single cell layer model was concentration dependent,and the transport volume increases with the increase of drug concentration.The apparent permeability of Andrographolide total Sulphonates and Potassium Sodium Dehydroandrographolide Succinate on the Calu-3 single cell layer model is larger.The permeability of the respiratory tract is higher and is more suitable for the development of lung inhalation preparation.The apparent permeability of Andrographolide Sodium Bisulfite and Potassium Dehydroandrographolide Succinate on the Caco-2 single cell layer model is larger.The mucous membrane of the digestive tract is more permeable and is more suitable for the development of oral administration.The efflux ratio of Potassium Sodium Dehydroandrographolide Succinate in Calu-3 single cell layer model and Andrographolide Sodium Bisulfite in Caco-2 single cell layer model are significantly lower when the P-glycoprotein inhibitor was added.So P-glycoprotein participates in the process of transport.Therefore,combined application of P-glycoprotein inhibitors can reduce efflux,promote absorption and increase permeability.