| Objective: To investigate the therapeutical effect of Penthorum Chinese Pursh extract of different concentration ethanol on alcoholic fatty liver in rats, and detecte the effects on serum lipid level, serum transaminase level and serum inflammatory factors level. Histopathological changes of liver were observed, and acquired the best concentration of Penthorum Chinese Pursh extract. Explore the possible mechanism of therapeutic effect on alcoholic fatty liver by Penthorum Chinese Pursh extract in rats. To provide experimental evidences for Penthorum Chinese Pursh were used to treat alcoholic fatty liver.Methods:(1) The main components of Penthorum Chinese Pursh were extracted by 35%, 55%, 75% and 95% alcohol.(2) 100 SD rats were divided into model group and control group randomly, model group rats were feeded with high fat diet containing iron and with 52% alcohol gavage for 8 weeks. The rats control drinking water also. So, the alcoholic fatty liver model was maded. The control group rats with common feed and free drinking.(3) The model group rats were divided into Penthorum Chinese Pursh tretment group and tiopronin tretment group, and were given extracts of different concentrations of ethanol Penthorum Chinese Pursh or tiopronin respectively.(4) During the treatment period, rats were observed on situations of spirit and behavior, etc. The change of liver were observed by visual inspection after killthe rats and the liver index was calculated.(5) The rats were fixated with 4%paraformaldehyde perfusion, and the livers were removed and fixed, paraffin-embedded, sections and hematoxylim-eosin staining. Histopathology changes of liver were observed under light microscopy; The serum transaminases level and serum lipid content were determined. Determination of the conten of malondialdehyde(MDA)and superoxide dismutase(SOD) in liver homogenate. Used enzyme-linked immunosorbent assay(ELISA) to detect serum levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), interleukin-6(IL-6)and nicotinamide adenine dinucleotide phosphate(NADPH).(6) The expression of P4502E1(CYP2E1), TNF-α, IL-1β, and IL-6 in liver tissue were detected by immunohistochemistry method(Envision method).Results:(1) The rats of model group appeared performance such as unresponsive, slow, drowsiness, sparse hair and dull, etc after 8 weeks which. high-fat diet containing iron and 52% alcohol gavage liquor, and the body weight of rats increased significantly, there was statistically significant(P<0.05) compared with the control group.(2) Liver index(liver weight / body weight): Compared with the control group, it was no significant difference(P>0.05) among the drug concentration treatment groups.(3) The macro changes of liver of rats were as follow. The livers of model group rats were gray soil, thick blunt edge, the volume increases, poor flexibility and fragility increases. The color, size and texture of liver of each treatment group were between the control group and model group.(4) The histopathology changes of liver of rats were asfollow. Hepatic cord and liver sinusoidal were blurred in model group rats under light microscope, and the lobule structures were disorder, vary sizes lipid droplets, spotty necrosis and inflammatory cell infiltration were seen visibly in liver cells. Group treated with Different concentrations of ethanol Penthorum Chinese Pursh extract show steatosis decrease, a small amount of inflammatory cell infiltration visible, the basic cells arranged neatly, lobular structure basically intact, and there is a small amount of spotty necrosis.(5) The effects of Penthorum Chinese Pursh extract on lipid metabolism in rats. Compared with the model group, 35%, 55%, 75% and 95% Penthorum Chinese Pursh extracts treatment group and tiopronin treatment group were significantly decreased serum triglyceride(TG), low density lipoprotein(LDL) and total cholesterol(TC) content(P<0.05), increased high-density lipoprotein(HDL) content(P<0.05) in rats. There was significantly difference while the concentration as 95% of Penthorum Chinese Pursh extracts treated group compared with the other treatment groups(P<0.05), but there was no significant difference in serum TG lowering effect with other treatment groups(P>0.05).(6) The effects of Penthorum Chinese Pursh extract to serum transaminases levels in rats. Compared with the model group, 35%, 55%, 75% and 95% Penthorum Chinese Pursh extracts treatment group and tiopronin t r e a t m e n t g r o u p w e r e s i g n i f i c a n t l y d e c r e a s e d s e r u m a l a n i n e aminotransferase(ALT)and aspartate aminotransferase(AST)content.(P<0.05), but between concentration as 95% of Penthorum Chinese Pursh extracts groupand tiopronin treatment group showed no significant difference(P>0.05).(7) The effects of Penthorum Chinese Pursh extract to liver homogenate MDA and SOD content in rats. Compared with the model group, 35%, 55%, 75% and 95% Penthorum Chinese Pursh extracts treatment group and tiopronin treatment group were significantly decreased the MDA content but increased SOD content in liver tissue(P<0.05); there was significantly difference while the concentration as 95% of Penthorum Chinese Pursh extracts treated group compared with the other treatment groups(P<0.05).(8) The effects of Penthorum Chinese Pursh extract to serum inflammatory cytokine levels in rats. Compared with the model group, 35%, 55%, 75% and 95% Penthorum Chinese Pursh extracts treatment group and tiopronin treatment group were significantly decreased serum NADPH, IL-1β, TNF-α and IL-6 levels(P<0.05); there was significantly difference while the concentration as 95% of Penthorum Chinese Pursh extracts treated group compared with the other treatment groups(P<0.05).(9) The effects of Penthorum Chinese Pursh extract to associated inflammatory factors expression in rat liver tissue. Compared with the control group, rats in the experimental group shew that the expression of cytochrome P4502E1, IL-1β, TNF-α and IL-6 were significantly enhanced, its gray value was significantly higher(P<0.05); the expression of the factors above-mentioned in each treatment group was significantly reduced; and compared in different concentrations of the treatment group, the expression of IL-1β and TNF-α in 95% Penthorum Chinese Pursh extracts group was lower than other treatmentgroups(P<0.05); and cytochrome P4502E1 and IL-6 expression was no significant difference(P>0.05 between the treatment groups).Conclusion: Different concentrations of Penthorum Chinese Pursh extracts have a better therapeutic effect to alcoholic fatty liver rats. But the effect of 95% Penthorum Chinese Pursh extracts is the best which reduce ALT, AST, TC, LDL and MDA, and increased HDL and SOD while reduce serum inflammatory factors associated compare with the other Penthorum Chinese Pursh extracts treatment groups. The possible mechanism are related to inhibite the activation of inflammatory cytokines, enhance the body’s antioxidant capacity, and remove the body’s oxygen free radicals. |
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