Pharmacodynamics And Pharmacokinetics Study On Resveratrol And Polydatin

Objective: In this study, human hepatoma carcinoma cell HepG2, human breast cancer cell MDA-MB-231 and human breast cancer cell MCF-7 were employed to investigate the inhibitory effects of resveratrol and polydatin, and to study the uptake and metabolism of these two drugs in two aspects-pharmacodynamics and pharmacokinetics. The purpose of this study was to provide some basic data for the application of resveratrol and polydatin.Methods:1. MTT assay was used to investigate the growth inhibitory effects of resveratrol and polydatin in HepG2, MDA-MB-231 and MCF-7 cells in different concentrations (15, 30, 50, 100, 150 and 300μmol/L).2. Flow cytometry was applied to evaluate the apoptosis of HepG2 cells induced by different concentrations(1, 30 and 300μmol/L)of resveratrol and polydatin.3. Resveratrol-glucuronide and polydatin-glucuronide were synthesized respectively.4. The uptake of resveratrol and polydatin by HepG2 and MDA-MB-231 cells was observed with fluorescent microscope. 5. HepG2 cells were incubated with 100μmol/L resveratrol and polydatin respectively for 15, 30, 60, 120, 180 mins, or were incubated with different concentrations (10, 20, 50, 100, 200μmol/L) of resveratrol and polydatin. Then LC/MS/MS(ESI + )was used to detect resveratrol, polydatin and their glucuronidated metabolites in HepG2 cell lysate in selected multiple reaction monitoring(MRM)mode.Results:1. Both resveratrol and polydatin inhibited the cell growth in a dose-dependent manner. At 48 hours, the inhibitory effects of 30, 50, 100μmol/L resveratrol on HepG2 cells were weaker than those of polydatin at the same concentrations. But the inhibitory effect of 300μmol/L resveratrol was stronger than that of polydatin. And at 72 hours, the inhibitory effects of resveratrol on HepG2 cells were stronger than those of polydatin both at 150 and 300μmol/L. In the dose range of 15-300μmol/L, resveratrol and polydatin could inhibit the proliferation of MDA-MB-231 cells in a dose-dependent manner. Compared with incubation for 48 h, the inhibitory effects were stronger after incubation for 72 h. Resveratrol were more effective than polydatin in the same concentrations. The inhibitory effects of resveratrol on MCF-7 cells were in a dose-dependent way. At 48 hours, polydatin could not significantly inhibit the proliferation of MCF-7 cells except for 300μmol/L polydatin. While, at 72 hours, the inhibitory effects of polydatin increased in a dose-dependent manner. Resveratrol were more effective than polydatin on MCF-7 cells in the same concentrations.2. Both resveratrol and polydatin at higher concentrations(300μmol/L) induced apoptosis of HepG2 cells significantly, and resveratrol was more effective.3. One substituted glucuronidated resveratrol and one substituted glucuronidated polydatin were synthesized respectively. 4. The fluorescent microscope image results indicated that both resveratrol and polydatin were quickly uptaken by HepG2 cells and MDA-MB-231 cells, and the amounts of the resveratrol and polydatin in cells increased with time increasing.5. Both resveratrol and polydatin were quickly uptaken by HepG2 cells, and resveratrol achieved its maximum concentration in HepG2 cells at the 30 min. The uptake of polydatin increased in a time-dependent way and was still unsaturated until 180 min. The amount of resveratrol uptaken by HepG2 cells was much more than that of polydatin at the same concentrations. The concentration of resveratrol at 30 min was 48.9μmol/L·mg protein, which was 38- fold higher than that of polydatin (1.3μmol/L·mg protein). The uptake of resveratrol and polydatin increased with the increase of the dose, and the uptake by the cells was still unsaturated at 200μmol/L. The amount of resveratrol absorbed by the cells was much higher than the amount of polydatin at the same concentration.Conclusions:1. Resveratrol and polydatin could significantly inhibit the growth of HepG2, MDA-MB-231 and MCF-7 cells in dose-dependent and time-dependent manner. The inhibitory effects of resveratrol were stronger than those of polydatin in the same concentrations(p<0.01). Compared with MCF-7 cells, MDA-MB-231 cells were more sensitive to resveratrol(p<0.05).2. Both resveratrol and polydatin at higher concentrations(300μmol/L)induced apoptosis of HepG2 cells significantly, and resveratrol was more effective at the same concentration(p<0.05).3. The LC/MS/MS method for simultaneously detection resveratrol, polydatin, one substituted glucuronidated resveratrol and one substituted glucuronidated polydatin, was proved to be accurate, sensitive and suitable for pharmacokinetic study.4. The uptake of resveratrol and polydatin by HepG2 cells increased in dose-dependent and time-dependent manner. The absorption velocity and the amount of resveratrol was much higher than the amount of polydatin(p<0.05) in HepG2 cells.5. From this study, it could be concluded that resveratrol was more potential for clinical application than polydatin.