Uptake Characterizations of Berberine, Palmatine and Jateorhizine in Rat Primary Hepatocytes / 中国药学杂志

ABSTRACT

OBJECTIVE: To evaluate the uptake characterizations of berberine, palmatine and jateorhizine through hepatic uptake model with rat primary hepatocytes. METHODS: Rat hepatocytes were isolated by Seglen's two-step method and cultured in suspension. The activity of transporters was assessed using rosuvastatin which is known as the substrate of the transporters. The tested drugs were incubated with rat hepatocytes at 4 or 37℃ for various periods of time and the drugs were determined using UPLC-MS/MS method to calculate the uptake amounts. The effect of cultural time and temperature on the hepatic uptake of these compounds were assessed. The kinetic parameters such as Km, Vmax and CLactive/CLuptake were calculated from concentration dependent uptake experiments. The effects of positive inhibitors on the uptake of berberine, palmatine and jateorhizine were also surveyed by pre-incubation of the inhibitors with the rat hepatocytes, followed by co-incubation with berberine, palmatine and jateorhizine. RESULTS: The hepatic uptake of berberine, palmatine and jateorhizine increased along with the increase of concentration at 37℃ while the change of hepatic uptake was very little at 4℃. The value of CLactive/CLuptake was 85.26%, 74.90% and 57.74% for berberine, palmatine and jateorhizine. Ibuprofen, digoxin, glycyrrhizin and prazosin which were the known inhibitors of transporters could remarkably reduce the hepatic uptake of berberine and palmatine, respectively. Glycyrrhizin and prazosin could reduce the uptake of jateorhizine. CONCLUSION: The transport of berberine, palmatine and jateorhizine into hepatocytes is mainly through an active process. Berberine and palmatine are the substrates of Oatp1a1, Oatp1a4, Oatp1b2 and Oct1, and the uptake of jateorhizine might be associated with Oatp1b2 and Oct1.