Reversing Effects Of β-elemene On Rats With Hepatic Fibrosis

AIM: To study the effect of zedoary rhizome extract (β-elemene) on the expression of TGF-β1,α-SMA ,Col-Ⅰ,MMP-2 and MMP-9mRNA in Rats with hepatic fibrosis.MATERIALS AND METHODS: The experimental model of Wistar male rats'hepatic fibrosis was induced by hypodermical injection of carbon tetrachloride (CCl4).β-elemene was administrated to the treatment group for 8 weeks with intraperitoneal injection (0.1ml/100g body weight) everyday. The samples were stained with hematoxylin and eosin for histopathological examination. Masson stain was used for liver fibrosis. Liver functions were measured by enzymatic kinetic analysis. Expressions ofα-SMA, TGF-β1 and collagen type I in the liver tissues were measured by SP-immunohistochemistry method. Contents of hydroxyproline in the liver tissues were tested by specimen alkaline hydrolysis. The ratio (V/B) of integrated optical density of MMP-2 and MMP-9 mRNA to that of bata-actin mRNA was calculated by semi quantitation reverse transcriptase polymerase chain reaction (RT-PCR). The relative expression quantity of MMP-2 and MMP-9 mRNA was represented by V/B.RESULTS: After eight weeks of treatment, the percentage areas of collagen fiber in normal, model, control and treatment groups were 1.22±0.24,7.47±0.81,5.57±0.78 and 4.33±0.48,respectively. The histological remission and collagen fiber diminish were significantly better in treatment group than in model group and control group (treatment group vs model group, P<0.01; treatment group vs control group, P<0.01). As shown in our experiment that the Serum ALT/AST/ALB activity (IU/L: mean±SD) were 254.88±70.18 vs 208.47±85.70, 414.42±138.11 vs 330.92±107.40, 26.54±1.59 vs 28.47±1.11, P<0.05, respectively, between model group and treatment group. Its also shown that the expression of HYP in liver of four groups were 0.162±0.048,0.410±0.145, 0.273±0.088, 0.186±0.120, respectively (treatment group vs model group, P<0.01).we clearly provided direct evidence that the expression of collagen type-I in the groups were 3.022±0.553,9.998±1.431,7.554 ±0.914 and 4.587±1.008,respectively, which also significant deviation compare with each other (treatment group vs model group, P<0.01; Treatment group vs control group, P<0.01). The expression ofα-SMA in liver tissues were 0.968±0.281, 5.605±1.315, 4.914±0.893 and 3.172±0.542, respectively (treatment group vs model group, P<0.01). The expression of TGF-β1 in liver of the groups were 1.026±0.208, 5.653±0.9, 4.702±0.627 and 2.868±0.554, respectively (Treatment group vs model group, P<0.01; Treatment group vs control group, P<0.01). The expression of MMP-2mRNA in liver tissues were 0.7124±0.0148, 0.8398±0.1068, 0.7876±0.1159 and 0.8211±0.0926, respectively (model group vs normal contral group, P<0.01). The expression of MMP-9mRNA in liver tissues were 0.5804±0.0145, 0.3268±0.2690, 0.3302±0.1685 and 0.3678±0.2577 respectively (model group vs normal contral group, P<0.01). The expression of TIMP-1mRNA in the model group increased significantly compared with normal contral group. (0.6799±0.1245, 1.7659±0.1459, 1.2293±0.0774 and 0.9294±0.0752, respectively,model group vs normal group P<0.01,treatment group vs model group P<0.01,treatment group vs control group P<0.01)The V/B of MMP-2mRNA which we measured in liver tissue of model group is more than normal control group and treatment group. (normal control group vs model group, P<0.01; treatment group vs model group, P<0.05), their V/B are 1.0124±0.1257, 1.9353±0.1447, 1.5885±0.2963 and 1.3205±0.3875 respectively. The V/B of MMP-9mRNA which we measured in liver tissue of the groups were 0.1684±0.0745, 0.5137±0.0918, 0.3833±0.1605 and 0.3178±0.1204 respectively (normal control group vs model group, P<0.01; treatment group vs model group, P<0.05)CONCLUSION:β-elemene can reverse the pathologic progression of fibrosis. The proposal that regression of liver fibrosis is mediared by inhibited activation of hepatic stellate cells ,down-regulated expression ofα-SMA,TGF-β1, collagen typeⅠ,enhanced expression of MMP-2mRNA and MMP-9mRNA and decresed sediment of extracellular matrix in the liver tissues.