Effect Of Ge Huang Antialcoholism Granules On Rats In ALD Model And The Study Of Its Mechanism

Objective: Through animal experiment, we set out to observe the efficacy of Ge Huang Antialcoholism Granules(GHAG) to treat alcoholic liver disease(ALD) rat model and the possible influence on oxidative stress and lipid metabolism, thus, figuring out the potential of this traditional Chinese medicine in ALD therapy and uncovering the underlying mechanism.Methods: 108 SD rat were randomly divided into 6 groups: normal group, model group, low dosage GHAG treatment group(low dosage group), middle dosage GHAG treatment group(middle dosage group), high dosage GHAG treatment group(high dosage group), metadoxine treatment group(control group). 18 rats were included in each group.ALD model establishment and drug intervention: Rats from model group, low dosage group, middle dosage group, high dosage group and control group were daily given 40% alcohol by gavage at a dosage of 1ml/100 g and supplemented with high fat dietary on the next day. On the contrary, rats from normal group were subjected to same volume of dd H2 O by gavage as well as normal dietary. Four weeks later, the low, middle and high dosage group of rats were administrated with GHAG at corresponding dosage. Metadoxine was used for control group. Drug intervention persisted for 8 weeks.The histological morphology of liver was assayed by HE staining. Serum AST, ALT, TG, TC was quantified by automatic biochemical analyzer. Real-time PCR was employed to analyze the RNA level of PPAR-α and P4502E1. For P4502E1, western blot was also used to determine the protein level. The liver tissue content of MDA, SOD and GSH-PX were determined by Thiobarbituric acid method, Xanthine oxidase method and ultraviolet spectrophotometry method, respectively. And the TG and TC level was quantified by colorimetric method.Results: 1.As indicated by HE staining, the liver from normal group rat showed normal structure with clear intercellular boundary and finely radially-orientated cell population toward portal tube. On the contrary, the model group presented obviously swelling liver cell with heterogeneously sized fat vacuole, obscure intercellular boundary, abnormally positioned nucleus, disordered hepatic cord. However, drug treatment in the high dosage group and control group rescued the liver structure to averagely normal level and the middle dosage group also showed improved histological morphology. While, the low dosage group show similar structure compared with model group.2.Compared with the normal group, model group rats showed significantly increased serum level of AST, ALT, TG, TC and liver content of TG, TC, MDA, as well as elevated P4502E1 expression both at m RNA and protein level(P<0.05). However, these can be rescued markedly by high dosage of GHAG(high dosage group) and metadoxine(P<0.05). The middle and low dosage of GHAG can also reversed above index but without statistical significance(P>0.05). Still, no statistically significant difference was observed between high dosage group and control group(P>0.05).3.Rats in model group demonstrated significantly decreased SOD and GSH-PX activity and PPAR- α m RNA expression(P<0.05). High dosage of GHAG(high dosage group) and metadoxine can significantly reverse the decrease of SOD, GSH-PX and PPAR-α m RNA(P<0.05). Although middle and low dosage of GHAG can also increase the above three indexes, the extent showed no statistical significance(P>0.05). Same as above, no statistically significant difference in respect to SOD, GSH-PX activity and PPAR-α m RNA level was observed between high dosage group and control group(P>0.05).Conclusion: 1. Model construction resulted in significant lipid accumulation in liver cell accompanied with increased liver weight and liver index, impaired liver physiological function, elevated blood lipid level. This was further confirmed by the observation of fat vacuole with HE staining.2. GHAG can decrease the accumulation of lipid in liver and blood lipid level, thus ameliorating liver function in ALD rat model. Our results indicated efficient therapeutic effect of GHAG for ALD.3. GHAG can lower the level of P4502E1 m RNA and protein and MDA level, increase the PPAR-α m RNA, SOD and GHS-PX level, the regulation of oxidative stress and lipid metabolism may be involved in the GHAG mediated therapy for ALD.