| PrefaceAlcohol - induced disease ( ALD) is a frequent liver disease in the western world. For decades, the incidence of ALD increased gradually in our country. Treatments of ALD are limited to supportive measures because the mechanisms involved in ALD are poorly understood. Currently, the studies of medicine against ALD are at the experimental stage in external. Internal scholars pay more attention to the Chinese herbal medicine in immune or chemical liver injures. The effects of Chinese herbal medicine to ALD are undocumented and there were no documents on the expression of basic fibroblast growth factor (bFGF) in alcoholic -induced disease by now.In our study, we investigated the effect on alcohol - induced disease of pre -using Kang Xian Fu Fang I ( KXI) in the rats. Mechanisms were studied if KXI could prevent alcohol - induced fibrosis. The expression of bFGF was detected by immunohistochemistry, which might provide information for deep understood the pathogenesis of ALD.Materials1. Experimental animal; 90 Wistar rats weighing between 160 and 200 g.2. Agent.- ethanol, Kang Xian Fu Fang I, radioactive immunoas-say agent box of HA and IV - C, antibody against bFGF, SABC im-munohistochemistric agent box.3. Apparatus: r - radioactive immunoassay count machine, transmission electron microscope, automated analyzer, computerized image analysis system, optical microscope, spectro photometer.Methods1. Alcoholic - Induced Hepatic Disease ModelRats were divided into three groups randomly. Alcohol - fed rats were infused ethanol intragastricly thrice a day for 12w. 0 - 4w, 400mL/L ethanol 8g? kg-1 · d-1;4 - 8w,500mL/L ethanol 9g? kg-1· d-1 ;8 - 12w,500mL/L ethanol 10g · kg-1· d-1. Medicine -treated rats were admission KXI (4ml · kg-1 · d-1) twice a day plus equal ethanol with Ethanol - fed rats for 12w. Control group received normal sodium (6ml? kg-1· d-1) instead of ethanol thrice a day for 12w.2. Histopathological AnalysisRats were killed at the end of 4w, 8w, 12w. A small sample of liver was fixed in formalin. Hematoxylin - eosin stain was used for light microscope. For evaluation of fibrosis, sections were stained with VG and analyzed using computerized image analysis. Liver ultra - thin sections were made to observe the changes of liver ultrastructure by e-lectron microscope.3. bFGF expressionThe expression of bFGF was detected by immunohistochemistry. Paraffin hepatic sections were stained with SABC method. The first antibody against bFGF was diluted in 1:50. For evaluation the expres-sion of bFGF, the rate of positive areas to hepatic areas was detected by Image Analysis.4. Hepatic function and serum HA and V - C measurements Serum hepatic enzymes were measured using an automated analyzer. Serum HA and IV - C were measured by radioactive immunoas-say.5. Liver MDA and SOD MeasurementsTBA method was used for liver MDA measurement. SOD was examined by hydroxylamine method.6. Statistical AnalysisAll data were expressed as means SD. The Statistical significance was determined by student' s t test.Results1. Serum hepatic enzymes and HA and IV - CSerum ALT, ALP and IV - C were both significantly increased after eight weeks ethanol feeding in alcohol - fed rats( P <0. 05). At the end of 12 weeks, ALT, ALP and IV - C reached the highest levels and serum HA was significantly higher in alcohol - fed rats than control and medicine - treated groups( P <0.01).2. Liver MDA and SODLiver MDA significantly increased at the fourth week in alcohol -fed group compared with control and medicine - treated groups ( P < 0.05). With time of ethanol exposure, liver MDA increased gradually. Ethanol admission caused an increase in liver SOD and there no significantly differences among the alcohol - fed and medicine - treated rats at the end of 4 weeks. However, SOD decreased gradually in the alcohol - fed rats, whereas values were higher continuously thanthe alcohol - fed group ( P < 0.05).3. HistopathologyHistological results w... |
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