Hepatoprotective Efficacy And Pharmacokinetic Study Of Antrodia Cinnamomea Dropping Pill

Objects:1.Antrodia cinnamomea fruiting bodies of water and alcohol extract preparation into dripping pills dosage forms, the establishment of dripping pills quality control standards, to determine the best Antrodia cinnamomea dripping pills (ACDP)prescription and completed can be prepared by technology research.2. Comparison of Antrodia cinnamomea fruiting bodies dripping pills and the powder of Antrodia cinnamomea fruiting bodies after oral administration in mice in vivo drug absorption pharmacokinetics between degree of blood concentration.3. Observation of Antrodia cinnamomea fruiting bodies of dropping pills on the intraperitoneal injection of APAP the rat liver function indexand changes in circumstances, as well as antioxidant enzymes suchas SOD and GPx activity performance, and its efficacy for the protection of the liver damage.Methods:1. Using solid dispersion technique, Antrodia cinnamomea fruiting bodies of water and alcohol extract preparation into Dripping Pill formulations, and experimental design to find the best prescription. Which the column were isolated and purified a known hepatoprotectivc the composition Antcin R, as the index composition analysis of ACDP ingredients. And use of liquid chromatography, Fourier transform infrared spectroscopy, electron microscopy and in vitro dissolution tests to establish the dripping pills quality control standards.2. By the serum pharmacokinetics testing, pharmacological studies of the oral bioavailability of ACDP, Intravenous and oral administration of two groups, according to themode of administration is divided into adult male surname Sprague-Dawley rats (230-280g) experiment, groups of six rats, a total of twelve. And jugular vein cannulation, as the blood taken pathway. ACDP dissolved in ethanol.a dose of50mg/kg, Antrodia cinnamomea powde at a dose of lg/kg and12hours after surgery, administration of0.2mL of blood taken from the jugular vein,to remove the protein, from the upper to clarifyliquid Antcin B content by HPLC analysis.3. Hepatoprotective pharmacological studios on the research group is divided into A. the normal control group:saline (IP)+ddH2O (PO) B. The negative control group (liver injury group):400mg APAP/kgbw (IP)+d. d. H20(PO) C. The control group-acetylcysteine (NAC) treatment control group):400mg APAP/kgbw(IP)+600mg of NAC/kgbw(PO) D. ACDP single the dose (1X) experimental group:400mg APAPAgbw(IP)+ACDP (20/kgbw)(PO) E. ACDP three times the dose(3X) experimental group:400mg of APAP/kgbw(IP)+ACDP (60/kgbw)(PO) F. ACDP five times the dose (5X) the experimental group:400mg of APAP/kgbw(IP)+ACDP (100/kgbw)(PO). A group of intraperitoneal injection of saline(0.6ml/30g bw formice.), The B-F group by intraperitoneal injection of400mg APAP/kg bw (0.6ml/30g bw for mice.)Twice a week (Monday and Thursday) in the samples were fed one hour before induction ofhcpatitis, all animals weekly administration of the test sample(d. d.1120or NAC),four hours after tail blood biochemical detection of liver function:GOT (AST), GPT (ALT). Finally, at the end of the eighth week, all the expense of laparotomy and liver specimens for pathological examination and do special staining of Masson’strichrome. In addition to the remaining liver were the detection of anti-oxidant glulathione (GSH), glutathione peroxidase (GSHPx) and glutathione reductase,(GSHRd), glutathione S-transferase (GST) of superoxide dismutase(SOD) and catalase enzyme activity.Results:1. According to the separation and purification of Antrodia cinnamomea extract and HPLC results. Established hepatoprotective the ingredients Antcin B, for the quality control standard and validation. Antrodia cinnamomea after the emulsification of water and alcohol extract of the solid matrix (PEG6000) melt into the solid dispersion solution using solid dispersion technology. The use of experimental design method to find the best conditions of ACDP is triterpenoid components:carrier:emulsifier=1:2.5:0.5. Established and completed ACDP of optimal conditions, quality standards and the preparation process. Uniform quality of the sample used in this study.2. The bioavailability pharmacokinetics experimental results obtained, the concentration in the blood of the rat body measured of obtaining Antcin B, ACDP0.0470μg/ml Antrodia cinnamomea powder0.0200μg/ml;reached the highest the blood concentration time ACDP0.50hr faster than powder0.75hr Antrodia cinnamomea.3. GOT and GPT index of the treatment groups, compared with negative control group. With the increase of ACDP dose, a dose-related reduce. GSH concentration than the negative control group was significantly improved; and a dose-related. GPx activity with increasing doses of ACDP, the gradual recovery;Mn-SOD activity, or of Cu/Zn-SOD activity were gradually increased with increasing Dripping Pills dose of Antrodia cinnamomea. Pathological observation:five times the dose (5X) ACDP under pseudo-lobule within the liver tissue formation and inflammatory cell infiltration from moderate into a small amount of the degree; the same time, the central vein andportal interval septum formation is less complete, show the degree of hepatic fibrosis is minor than the negative control group.Conclusion:1. separation and the establishment of the hepatoprotective ingredients Antcin B, quality control standards.2. To determine the best ACDP prescription, and completed research can be prepared by process.3. ACDP for the liver cells have a protective effect.4. ACDP than the powder to absorb quickly, a higher absorption rate. Dripping Pills can increase the the triterpenoids oral, absorption, the role of rapid inc.reasebioavailability.5. ACDP can make because of the APAP damage of liver cell mitochondria and the reticulo endothelial system of organizations back to normal,for liver function or antioxidant enzymes reply. For the liver cells ACDP have a protective effect of scientific evidence.