Study On The Anti-HBV Material Foundation Of Zijingantai

Zijingantai constitutes with Swertia punicea Hemsl and Jineijin , both them regard Swertia punicea Hemsl as the main medicine. In order to elucidate the material foundation of Zijingantai on treatment hepatitis, for the first time, we use the bioassay -guided fractionation of Swertia punicea Hemsl and Jineij in to find active fractions for anti-HBV and then clarify active constituents of Swertia punicea Hemsl, establish a method for determination contents of five active constituents in Swertia punicea Hemsl and other plants in Gentianaceae and establish HPLC-FPS of Swertia punicea Hemsl.Alcoholic extract of Swertia punicea Hemsl (SP) was dissolved in water and fractionated sequentially with Petroleum ether,ether and EtoAc. The fractions were concentrated to yielded petroleum ether fraction(F₁-SP),ether fraction(F₂-SP) , EtoAc fraction(F₃-SP) and remainder of water fraction(F₄-SP). F₄'-SP was collected by the AB-8 resin from F₄-SP. By the CPE,the cytotoxic effect of SP, F₂-SP , F₃-SP, F₄-SP and F₄'-SP were assayed . Anti-HBV activity of each fraction was evaluated by using radio-immunoassay on HBsAg and HBeAg in the cultured hepatocarinoma cell line HepG₂(2215 cells), which was transferred by hepatitis B virus(HBV) DNA. F4-SP and F₄'-SP show the lest cytotoxic effect, their TC₅₀ are more than 2000 μg/ml. F₃-SP exhibits anti-HBV activity in vitro, supprssing HBsAg and HBeAg , IC₅₀ are 58 μg/ml, 100μg/ml ;SI are 12.2,7.1, respectively.The four compounds were isolated from F₄'-SP. By assay of physical, chemical and spectroscopic data of UV, IR, ¹HNMR,¹³CNMR ,HMQC,HMBC and MS, the structures of these four compounds were determinated as mangiferin( â… ), swertiamarin (â…¡), wertianolin(â…¢) and glucose (â…£ ). The five compounds were isolated from F3-SP .The structures of them were identified as swertianin(â…¤), 1,7-di- hydroxy-3,8-dimetho xyxanthone(â…¥), bellidifolin(â…¦), 5-o-p-D-glucopyranosyl-l ,8-dihydroxy-3-metho -xyxanthone(â…§) and oleanolic acid(â…¨). Compound â…£, â…¤ and â…§ were obtained from Swertia punicea Hemsl for the first time. Compound II, VI, VII and IX suppress HBsAg expression of 2215 cells, their IC₅₀ are 432μ g/ml, 30...