Studies On The Establishment Of HPLC Fingerprint Of Agrimonia Pilosa Ledeb And Its Antitumor Mechanisms

Agrimonia pilosa ledeb, which is also called agrimony,is the dry aerial parts of agrimony rosaceae.lt takes actions as hemostasis with astringents, stopping dysentery, reinforcing deficiency and anthelminthic. Studies in modern medicine showed that it can be used in the treatment of lung cancer, liver cancer, gastric cancer and intestinal cancer in clinic. We have built up the HPLC fingerprint of Agrimonia pilosa ledeb and studied the antitumor mechanism of Agrimonia pilosa ledeb's water extraction.We apply HPLC to compare the fingerprints of Agrimonia Pilosa ledeb collected from different producing areas. Chromatographic system: Waters 616 high pressure continuous nonpulsatile flow pump,717 autosampler,996 diode array detector,20 1 0 work station. Chromatographic condition: Hypersil ODS column was used, with mixtures of 1% phosphoric acid and water as a mobile phase in a gradient mode. Detective wavelength was set at 280nm.Sample size is 10 u L. Flow rate is 1 mL min-1 .Mobile phase and gradient procedure acetonitrile : 1% phosphoric acid and water(time:A%): (0-0) - (20-20) - (30-20) - (31-40) - (47-60) - (55-90) - (56-100) - (70-100)o The relative retention time of each characteristic absorption band in the fingerprint is within the range of the mean relative retention time by + 1 , RSD of relative peak area are all less than 5%. Chromatographic condition showed thatAgrimonia from 8 different origins all have the same seven chromatographic peaks, which can be called common peaks. The set-up fingerprint have good reproducibility, high precision and stability .It can be used in the identification and quality control of Agrimonia .The Interference of cell cycle and proliferation of BGC-803 and Hela cell by water extraction of Agrimonia (drugs) is studied by electronic microscopy and flowcytometry respectively, and the IL-12 and TGF-B are also detected by ELISA in order to explore the pharmacological mechanism of Agrimonia. The experimental results demonstrate:the results by electronic micorscopy and opticmicroscopy all support that the water extraction of Agrimonia can inhibit the proliferation of BGC-803 and Hela in vitro,there are quite dictinct change happened after the treatment in cellphology and cell ultraconstitution. The inhibition ratio increased greatly as the concentration of water extaction of Agrimonia is raised.When the drug concentration is 100 u gmL-1,the inhibition ratio of BGC-803 and Hela are 61.9% and 67.9%,respectively. The results of flow cytometry proved that drugs could interfere the cell cycle and inhibit the transforming of cells from G1 + G0 phase to G2+M phase,called as cell cycle special drugs.The results of ELISA confirmed that drugs could increase IL-12 expressing and decrease TGF-B expressing,this can promote the immune attack and recognition in vivo, it might be one of the anti-tumorpharmacology mechanisms of drugs.