Screening the Lipid-lowering Active Ingredients in Rhubarb by the Hepatocyte Affinity Chromatography in Vitro / 世界科学技术-中医药现代化

ABSTRACT

Objective: To screen and analysis of antihepatic steatosis active components and cell metabolism withinrhubarb anthraquinones in vitro. Methods: Lipid-lowering activity of liver cell affinity components in rhubarbanthraquinones was evaluated by hepatic steatosis in vitro model. Hepatocyte-specific affinity compositions werescreened by hepatocyte affinity chromatography. Hep G2 cells were incubated with rhubarb anthraquinones in vitro andwere detected by HPLC and HPLC-Q-TOF-MS method. Results: Compared with normal group, the intracellular TGcontent of model group were significantly increased (P＜0.05) . Respectively treated with different concentrations ofrhubarb anthraquinones, the intracellular TG content were significantly decreased in a dose dependent manner (P＜0.05) in comparison with model group. Compared with rhubarb solution, there are mainly four hepatocyte-specific affinitycompositions of aloe-emodin, emodin, chrysophanol and physcion, except for rhein. Conclusion: The lipid-lowering experiment in vitro verified that five rhubarb anthraquinones have significant lipid-lowering activity. This research established a biochromatography method to screen out four hepatocyte-specific affinity compositions of aloe-emodin, emodin, chrysophanol and physcion from rhubarb anthraquinones, while the conjugation sites of rhein were ambiguous.Combined hepatocyte affinity chromatography with hepatic steatosis in vitro model, it could evaluate and screen lipidlowering active ingredient in vitro and cell metabolism of Chinese traditional medicine for the drug development of lipidlowering rhubarb anthraquinones.