The Antifibrotic Compounds Research Of Fuzheng Huayu Formula

Being conducted the strictly controlled random trials, the clinical effects of complex formulations of Traditional Chinese Medicine (TCM) have gained a lot proofs in Evidence-Based Medicine (EBM) thus to be approved by the international medical field. And the mechanisms of these effects have also been illustrated a lot. However, the formulations are so complicated that the chemicals responsible for their clinical affects as well as the metabolisms of these chemicals are not very clearly, which have drag down the modernization of them.In this thesis, we studied the antifibrotic effects in vivo of the Fuzheng Huayu formula (FZHY). The effective components into blood were detected by the HPLC/ESI-MS technique. Besides, the drug activity in vitro was analyzed by the High Content Screening (HCS).The rats were administrated with FZHY, and the serum from the rats was analyzed by HPLC/ESI-MS to study the effective into-blood components of the formula. Human stellate cells (LX-2) were stimulated with fetal bovine serum (FBS), and treated by the water-soluble into-blood component. The cell proliferation was evaluated by the MTT method. The human hepatic cells (HL-7702) apoptosis were induced by the TNF-a/Act D and interfered by the water-soluble into-blood components. The apotosis was detected by the relative kit of HCS.The results were as follows:1. Twenty-three components were identified in the serum of rats administrated with the fomulation. Among them, there were eight water-soluble compounds from Radix Salviae Miltiorrhizae such as Salvianolic acid B, Danshensu, Caffeic acid and Rosmarinic acid, et al; four fat-soluble compounds of salvia such as Cryptotanshinone, Tanshinone V, at al; eight lignans compounds such as Schisandrin, Schisandrol B and Schisantherin A, at al; and another two metabolites and amygdalin.2. The in vitro study of six water-soluble compounds from Radix Salviae Miltiorrhizae, including Salvianolic acid B, Danshensu, Caffeic acid, Rosmarinic acid, Protocatechuic acid and Protocatechuic aldehyde,showed that Salvianolic acid B, Danshensu, and Caffeic acid could inhibit the proliferation of LX-2 cells. And all of the compounds could inhibit the cell apoptosis of HL-7720 cells. Specially, Rosmarinic acid has the most toxic to the LX-2 cells but no toxic to the HL-7720 cells.In conclusion, Salvianolic acid B, Danshensu, Caffeic acid and Rosmarinic acid are a part of the effective components of Fuzheng Huayu formula;The in vitro activities of some of these components in blood were detected by HPLC/ESI-MS and HCS,this method is a simple, fast and effective way.