15 1 Vol. 15 No. 1 2002 2 Journal of Medical Postgraduates Feb. 2002 1, 2, 2, 1, 1, 1 1, (11, 200001 ; 21, 200070) 13 : :, (LBP) : ( 5 % 40 % 40 %, 29 ),25 LBP (250 mg/ kg 1 000 mg/ kg) :,,LBP (1 000 mg/ kg) : LBP : ; ; ; : R575129 : A : 100828199 (2002) 0120013204 Ξ Morphological observation of protective effects of lycium barbarum polysaccharide on experimental alcoholic liver disease in rats SONG Yu2lin 1, ZENG Min2de 2, LU Lun2gen 2, FAN Zhu2ping 1, MAO Yi2min 1, YANG Wen2zhuo 1, CAO Ai2ping 1 (1. Research Institute of Digestive Diseases of Shanghai, Renji Hospital, Shanghai Second Medical University, Shanghai 200001, China ; 2. The Centre Hospital of Zhabei Region, Shanghai 200070, China) Abstract : Objectives :To induce a relatively simple and feasible rat model of alcoholic liver disease(ald) and to investigate the protective effect of lycium barbarum polysaccharide(lbp) on it. Methods :Rat model of alcoholic liver disease was induced by receiving ethanol drinking water as the sole source of liquid. The ethanol concentration was increased from 5 % (ethanol/ water :v/ v) to a final concentration of 40 % for 29 weeks. After 25 weeks of treatment, survived rats were divided into 3 groups at random, i. e. ethanol2treated group, ethanol2treated plus two doses of LBP2treated groups (250 mg/ kg and 1 000 mg/ kg, respectively). Liver pathological changes under light and electron microscope were observed. Results :A varied degree of hepatic steatosis and hepatitis were induced. Electron microscopy revealed swelling or vacuolization of mito2 chondria in a part of hepatocytes, a marked proliferation of collagen fibrils in hepatic sinusoid was also ob2 served. Compared with the model group, LBP improved the pathological changes. The effects of 1 000 mg/ kg of LBP on ALD were better than that of 250 mg/ kg of LBP. Conclusions : The result indicated that LBP could improve morphological changes of mitochondria, alleviate steatosis and necroinflammatory changes in al2 coholic liver disease in rats. Key words : Lycium barbarum ; Alcoholic liver disease ; Rat ; Pathology 0 French Lieber2Decarli, 1995, Keegan, Tsukamoto2 1 Ξ : 2001208213 : (19662),
14 2002 2 15, ( / 100), 11312 H2E 10 % H2E 7, + 4 + 14 Knodell 8, 2 (lycium barbarum polysaccharide, LBP) 1,9 3,4 11313 1 5,6 mm 3 2. 5 % 24 h,0. 1 mol/ L 3,1 % LBP,HITACHI H - 500 1 111 11111 Wistar ( ) 75, (6112) ( Kruskal2Wallis ) 130150 g, 11112 2 LBP 211 ( :00918) 11 12 1 6, 112 ( P < 0. 000 1) 11211 75 36, Wistar 1 (15 ) (60 ) Keegan 1, 2 4 8 23 25 212 1 () 12,25 18 (250 mg/ kg) (1 000 mg/ kg) 0. 5 ml/ 100 g 5 % 20 %LBP ( ), 1, 4, 4 0. 5 ml/ 100 g, 1 114 x s, SAS (6. 12), Dunnett2t SAS 60 %, (12. 13 1. 27) g/ (kg d) 1 ( x s) Table 1 Body weight and the liver/ body weight ratio (g) 10 347. 40 31. 00 2. 63 0. 13 6 167. 33 34. 53 3 3. 41 0. 32 3 LBP(250 mg/ kg) 6 168. 67 23. 03 3 2. 82 0. 15 LBP(1 000 mg/ kg) 6 176. 00 18. 50 3 2. 71 0. 12 3, P < 0. 05 ;, P < 0. 05 11212 0. 1 g/ kg, 213 21311 (10 % ) (4 ; 2. 5 % 0. 1 mol/ L ),, - 70 113 21312 11311, ;,
1, 15 LBP, 13 (1, (2 2) 1),, () 2 3 2 Table 2 Pathological changes in the differential experiment groups - + 2 + 3 + 4 + 10 1 9 10 0 1 2 3 4 5 6 7 8 6 1 1 3 1 3 4 1 1 3 3 LBP(250 mg/ kg) 6 6 2 3 1 3 3 3 LBP(1 000 mg/ kg) 6 6 6, 3 P < 0. 01, 3 3 P < 0. 001, 3 3 3 P < 0. 01 ;, P < 0. 01 3 Table 3 Total pathological score in the differential experimental group 0 1 2 3 4 5 6 7 8 9 10 1 9 6 2 2 1 1 3 LBP(250 mg/ kg) 6 2 3 1 3 3 LBP(1 000 mg/ kg) 6 6, 3 P < 0. 01, 3 3 P < 0. 01 ;, P < 0. 01, P < 0. 05 21313, 3 ( 7 000) Figure 3 A few small lipid droplets appearing in hepatocyte of (3) a control rat (TEM 7 000) (4), (5), (6) 3 Keegan 1 4, ( 5 000), Figure 4 Hepatocyte from a rat fed ethanol showing many lipid droplets and a few organellae(tem 5 000)
16 2002 2 15 5 (,, 8 000) ; Figrue 5 Marked collagen fibrils appearing in hepatic sinusoid of a rat fed ethanol (TEM 8 000) 6 LBP,1993,7(4) :293. ( 8 000) Figure 6 Nepatocyte from a rat fed high dose of LBP show a few 6. lipid droplets and many organellae(tem 8 000), () H2E,,, TG TCh LDL2ch TG TCh, LBP CCl 4 (ALT) 5 6,LBP, LBP, 10,LBP 6,11 : 1 Keengan A,Martini R,Batey R. Ethanol2related liver injury in the rat : a model of steatosis,inflammation and pericentral fibrosis J Hepa 2 tology,1995,23 :5912600. 2 Hossein Sadarzadeh SM,Nanji AA. The 212aminosteroid 162desmethyl tiriliazad mesylate prevents necroinflammatory changes in experimental alcoholic liver diseasej Pharmacology Experimental Therapeutics, 1998,284 (1) :4062412. 3,.,1998,4(5) :54256. 4,,.,1997,19(4) :4152418. 5,,.,1989,3(3) :1632168. 7 Nanji AA,Jokelainen K, Rahemtulla A, et al. Activation of nuclear factor kappa B and cytokine imbalance in experimental alcoholic liver disease in the rat Hepatology,1999,30 (3) :9342943. 8 Knodell RG,Ishak KG,Black WC, et al. Formulation and application of numerical scoring system for assessing histological activity in asymp2 tomatic chronic active hepatitis Hepatology,1981,1 :431 2435. 9 Yin M,Wheeler MD, Kono H, et al. Essential role of tumor necrosis factorin alcohol2induced liver injury in mice Gstroenterology, 1999,117 (4) :9422952. 10 Rouach H,Fataccioli V,Gentil M, et al. Effect of chronic ethanol feed2 ing on lipid peroxidation and protein oxidation in relation to liver pathology Hepatology,1997,25 (2) :3512355. 11.,1993,18(2) :1102112. ( )
1,: X NF2 B ( 3 ) 1 pcdna 3. 12HBX HBV X mrna ( 400) Figure 1 HBV x mrna in hepatocellular carcinoma cell transfect2 ed with eukaryotic expression vetoc pcdna 3. 12HBX in situ hybridization ( 400) 4 TUNEL ALLN ( 100) Figure 4 Apoptotic hepatocellular carcinoma cell detected by TUNEL method ( 100) 5 ALLN ( 800) Figure 5 Apoptotic hepatocellular carcinoma cells detected by laser 2 NF2 B HBV x confocal microscope ( 800) ( 400),: Figure 2 Expression of NF2 B in hepatocellular carcinoma cell ( 15 ) transfected with HBV x gene. Immuno2fluorence staining ( 400) 3 ALLN NF2 B ( 400) Figure 3 Expression of NF2 B in hepatocellular carcinoma cell treated with ALLN. Immunohistochemical staining ( 400) 1, (H2E 300) Figrue 1 Liver from a rat fed ethanol showing marked steatosis prodominantly foam degeneration(h2e 300)
2 2 LBP 13 (H2E 300) Figure 2 Liver from a rat fed high dose of LBP showing steatosis in 13 layers of hepatocytes aroud the central vein(h2e 300) 3 (RTE11, 200) Figure 3 Levels of mucins expression of submucosal gland cells in2 creased in chronic bronchitis rats (RTE11 stain, 200),: ( 69 ) (H2E, 200) 1, Figure 4 Tracheal epithelia almost integrity, submucoal glands hy2, (H2E, 200) perplasia and hypertrophy, inflammatory cell infiltration Figure 1 Tracheal epithelia damage, submucosal glands hyperpla2 decreased in the dexamethasone group (H2E, 200) sia and hypertrophy, inflammatory cell infiltration in chronic bronchitis rats (H2E, 200) 4 2, 5 (RTE11, 200) (RTE11, 200) Figure 2 Tracheal goblet cells hyperplasia, levels of mucins ex2 Figure 5 Tracheal goblet cells hyperplasia, levels of mucins ex2 pression of epithelia increased in chronic bronchitis rats pression of epithelia decreased in the dexamethasone (RTE11 stain, 200) group (RTE11 stain, 200)