Study Of The Hepatoprotective Effect Of Extracts Of Sambucus

Sambucus Chinensis L. is a native perennial herb distributed throughout China. All parts of the plant can be used in traditional Chinese medicine (TCM ) as Luying. Luying is one of the important folk medicines in TCM. It has been used to treat hepatitis over a very long period of time and has produced quite a good effect.The effect of therapeutic hepatitis on Luying was studied in this paper. Luying extract by different extracting methods were prepared. Effects of Luying extracts of different processes on ALT and AST in acute hepatic injury mice were studied. As a result, 75% alcohol was selected as the optimal extraction reagent. The fractions of various polarities fractionated and purified with macroporous resin from 75% alcohol extract were further investigated, the fraction eluted from the macroporous resin with 30% alcohol(LY) were found to be primarily active. Then we primarily investigate the pharmacodynamics of against hepatitis of the LY .It showed significantly protective effects on mice or rats acute hepatic injury induced by CCU, D-GalN or Cona. On the other hand, it showed significantly improving the ability of nonspecific immunity of mice. In a word,the LY showed significantly protective effects against many kinds of chemical and immunological liver injuries, what's more, it has the effects improving the ability of nonspecific immunity of mice.The ursolic acid that was taken orally by mice or rats didn't show protective effects on mice or rats acute hepatic injury induced by CCl₄ or D-GalN, but it was reported by the literature that the ursolic acid that was taken orally has well effect of therapeutic hepatitis. So, a rapid, sensitive, and accurate liquid chromatography-mass spectrometry (LC-MS) method for the determination of ursolic acid in rat plasma was developed and validated. Plasma samples taken from rats that had received Luying extract orally were acidified with acetic acid and then extracted with a mixture of hexane-dichloromethane-2-propanol (20:10:1, v/v/v). Atmospheric pressure chemical ionization was operated in negative-ion mode. Using selected ion-monitoring mode, the deprotonated molecules [M-H]^- at m/z 455 and 469 were used to quantify ursolic acid and glycyrrhetic acid (internal standard), respectively. The assay was shown to be linear over the range of 10～1000 ng·mL^-1 (r≥0.9960) with a lower limit of quantification of 10 ng·mL^-1. The method was shown to be reproducible and reliable with intraday precision below 7.8%, interday precision below 8.1%, accuracy within±4.3%, and mean extraction recovery excess of 83.6%, which were all calculated from the blank plasma sample spiked with ursolic acid at three concentrations of 20, 200, and 800 ng·mL^-1. The LC-MS method has been successfully applied to pharmacokinetic studies of ursolic acid after oral administration of Luying ethanolic extract to rats. The main pharmacokinetic parameters were: t_1／2, 4.3h; C_max, 294.8 ng·mL^-1; and t_max, 1.0 h, respectively.We followed to go deep into studing the pharmacodynamics of against hepatitis of the LY and Luyingkeli (LYKL), what's more, there is a contrast between LY and LYKL. From the results, LY and LYKL could prevent liver from acute damage induced by CCl₄,D-Gal,ConA and TAA. They decreased the level of ALT and AST of blood serum in acute hepatic mice, and LY is well displayed the effects in dose-dependent manner. At the same time, LY and LYKL significantly decreased TG,MDA and GSH of liver tissue, and increased the level of T-AOC,T-SOD and Ca²⁺-ATPase of liver tissue of mice acute hepatic injury induced by CCl₄. On the other hand, LY and LYKL significantly decreased ALP,TBIL,TG and NEFA in serum, and increased the content of Glu in serum and glycogen and the activities of Na⁺-K⁺-ATPase,Ca²⁺-Mg²⁺-ATPase,Mg²⁺-ATPase,Ca²⁺-ATPase of liver tissue of mice acute hepatic injury induced by D-Gal. LY could increased the activities of SDH of liver tissue of mice acute hepatic injury induced by D-Gal, too. And the effect of LY is obviously better than LYKL.In chronic hepatic injured experiments, LY and LYKL could checkmate hypohepatia, and the effect of LY is obviously better than LYKL. At the same time, LY could obviously induced death rate, but LYKL not. LY and LYKL also certainly lowered hepatic cirrhosis rat the serum LN,HA,PIIIP contents and the liver homogenate HYP contents. From the pathologic section, compared with model group, LY and LYKL differently certainly improved the rat hepatic phathology. Rats hepatic cell necrosis, lipid degeneration, liver fibrosis were all abated. What's more, LY and LYKL could increase the ablity against lipoperoxidation of liver tisse. LY could increase the activities of Ca²⁺-ATPase of liver tissue, too. on the side, they showed significantly protective effects on the homogeneous immunological liver injury models of the cavy.In receding jaudice and normalizing gallbladder experiments, LY and LYKL had no obvious effect.In immunology experiments, LY and LYKL enhanced phagocyte function and accelarated carbon clearance in immunodepression mice induced by Hydrocortisone(HY). At the same time LY and LYKL stimulated antibody forming and increased hymolytic HC₅₀ in hypoimmunity mice induced by Cyclophosphamide(CTX). In lymphocyte of foetus proliferation experiments, LY and LYKL accelerate lymphocyte immunity. In lymphocyte of foetus proliferation experiments by LPS and ConA stimulated, LY showed certainly restrain effects, but LYKL had no obvious effect.