Isolation And Purification Of XieZhuoChuBidecoction Saponins By Macroporous Resin And The Study About Its Influence On GLUT9and XOD

Objects:1. To determine the contents of total saponins of the XieZhuo ChuBi decotion and obtain high-purity total saponins of XieZhuoChuBi decotion through screening the resins and the study of the best separation conditions;2. To study on whether the total saponins would has any influence on XOD;3. To obtain the perfect time and the levels of total saponins extracted from Xiezhuochubi decoction and study what effect it will has on GLUT9.Methods:1. To determine the contents of total saponins by UV spectrophotometry with456nm; and adopt the static adsorption-desorption to select the perfect macroporous resin, study the factors affecting the separation according to the content of total saponin;2. Uesd the XO-XOD reaction system, determine the absorb rate by uric kit, detecting the effect of total saponins on XOD;3. Determine the perfect time and the levels of total saponins in Xiezhuochubi decoction on the mouse renal tubular epithelia cells by MTT, and compare the genetic expression of GLUT9by RT-PCR. Results:1. The vanillin-acetic acid-perchloric acid system is the proper analyzing method of the total saponins in the XieZhuo ChuBi decotion, the gross of total saponins in the XieZhuo ChuBi decotion is10.28%; methodology examination reveales that the relationship between absorbency and mass concentration is linear, and the linear range is0.0426-0.2982mg·ml-1, r=0.9996, recovery rate is97.51%; The macroporous resin D-101not only possesses the strongest absorption ability, but also has an easy desorption property. The optimum absorption conditions were as follows:the loading volume is1.5BV, absorption rate is1.0BV·h-1and30%ethanol can elute most of the saponins with the volume of5BV, and the content of the XieZhuoChuBi decotion’s saponins is up to80%;2. The saponins can inhibit the XOD’s activity with the IC50at29.37μg;3. Total saponins extracted from Xiezhuochubi decoction wouLd not inhibit the RTECs in the range from50μg·mL-1to500μg·ml-1, and it also have the influence on the genetic expression of GLUT9.Conclusion:1. Total saponins can be extracted from Xiezhuochubi by macroporous resin;2. With the dosage of29.37μg, saponins can inhibit50%XOD’s activity.3. The total saponins in Xiezhuochubi decoction can lower the genetic expression of GLUT9.