ABSTRACT
PURPOSE: The degradation of the extracellular matrix has been shown to play an important role in the treatment of hepatic cirrhosis. In this study, the effect of thalidomide on the degradation of extracellular matrix was evaluated in a rat model of hepatic cirrhosis. MATERIALS AND METHODS: Cirrhosis was induced in Wistar rats by intraperitoneal injection of carbon tetrachloride (CCl4) three times weekly for 8 weeks. Then CCl4 was discontinued and thalidomide (100 mg/kg) or its vehicle was administered daily by gavage for 6 weeks. Serum hyaluronic acid, laminin, procollagen type III, and collagen type IV were examined by using a radioimmunoassay. Matrix metalloproteinase-13 (MMP-13), tissue inhibitor of metalloproteinase-1 (TIMP-1), and alpha-smooth muscle actin (alpha-SMA) protein in the liver, transforming growth factor beta1 (TGF-beta1) protein in cytoplasm by using immunohistochemistry and Western blot analysis, and MMP-13, TIMP-1, and TGF-beta1 mRNA levels in the liver were studied using reverse transcriptase polymerase chain reaction. RESULTS: Liver histopathology was significantly better in rats given thalidomide than in the untreated model group. The levels of TIMP-1 and TGF-beta1 mRNA and protein expressions were decreased significantly and MMP-13 mRNA and protein in the liver were significantly elevated in the thalidomide-treated group. CONCLUSION: Thalidomide may exert its effects on the regulation of MMP-13 and TIMP-1 via inhibition of the TGF-beta1 signaling pathway, which enhances the degradation of extracellular matrix and accelerates the regression of hepatic cirrhosis in rats.
Subject(s)
Animals , Male , Rats , Actins , Carbon Tetrachloride/toxicity , Collagen Type III/metabolism , Down-Regulation , Extracellular Matrix/metabolism , Immunohistochemistry , Immunosuppressive Agents/pharmacology , Liver Cirrhosis, Experimental/chemically induced , RNA, Messenger/analysis , Rats, Wistar , Thalidomide/pharmacology , Tissue Inhibitor of Metalloproteinase-1/biosynthesis , Transcription Factor RelA/biosynthesis , Transforming Growth Factor beta1/biosynthesis , Transforming Growth Factors/metabolismABSTRACT
OBJECTIVE:To provide reference for the establishment of quality standard for Fresh Gastrodia elata lyophilized powder. METHODS:TLC was conducted for the qualitative identification of G. elata in preparation,and HPLC was conducted for the content determination of gastrodin in preparation. The column was Diamonsil C18 with mobile phase of acetonitrile-0.05% phos-phoric acid(3∶97,V/V)at a flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 25 ℃ and vol-ume injection was 10 μl. RESULTS:TLC pots of Fresh G. elata lyophilized powder were clear and well-separated. The linear of gastrodin was 2.9-14.5 μg/ml(r=0.999 9);RSDs of precision,stability and reproducibility tests were no more than 1.00%;recov-ery was 98.07%-102.70%(RSD=1.74%,n=6). CONCLUSIONS:The method is simple,accurate and reproducible,and suitable for the quality control of Fresh G. elata lyophilized powder.
ABSTRACT
OBJECTIVE:To establish the quality standard for Gastrodia elata hency-fired tablet. METHODS:TLC was used to identify the gastrodin and benzyl alcohol and determine the content of moisture,ash and extract;HPLC was used to determine the content of gastrodin and benzyl alcohol. Column was Diamonsil C18 with mobile phase of acetonitrile-0.05% phosphate(3:97,V/V) at flow rate of 1.0 ml/min,detection wavelength was 220 nm,column temperature was 25 ℃ and volume injection was 10 μl. RE-SULTS:The TLC of gastrodin and benzyl alcohol showed clear spots and good separation. The moisture content40.0%. Linear range of gastrodin was 25.2-126.0,12.7-63.5 μg/ml(r=0.999 9),respectively;RSDs of precision,reproducibility and stability tests were lower than 2.0%;recovery was 99.49%-102.40%(RSD=1.09%,n=6), 98.75-102.63%(RSD=1.53,n=6),respectively. CONCLUSIONS:The method is simple and good reproducibility,and can be used for the quality control of Gastrodia elata hency-fired tablet.