| Objective:Acute hepatic failure (AHF) is a clinical syndrome from a variety of causes resulting from rapid loss in hepatocyte function, typically associated with coagulopathy, encephalopathy and digestive system which eventually can lead to multiple organ failure and death. AHF is one of the detrimental disease which hit severely, rapid developing, poor prognosis and its mortality rates can reach70%-80%. It’s very important to use Chinese drugs to protect suvrival hepatic cells and make liver rgeneration. The complexity of AHF clinical presentations makes it difficult to make treatment based on syndrome differentiation. But in our viewpoint, AHF belongs to the acme stage of liver disease, its main etiology are toxicity, blood stasis and Qi deficiency, the treatment of it must get rid of the evil and strengthen body resistance. AHF belongs to "acute jaundice " of traditional Chinese medicine(TCM), there is lack of effective drugs for AHF treatment, currently. According to the long-term clinical experience of liver disease of Guangdong province key subject of integrated Chinese and Western medicine in Nanfang Hospital, we made Sanhuangyinchi Decoction(SHYCD) to treat it. In this study, the rats were treated with D-GalN plus LPS to induce AHF.By obsevring the change of the indexes of SOD, MDA and Caspase3, the effects and probable mechanisms of SHYCD on rats with AHF has been explored.Methods:1. MedicineComposition of Sanhuangyinchi Decoction:Rheum officinale, Rhizoma Curcumae Longae,Astragalus mongholicus, Artemisia capillaries Thunb, Paeonia veitchii Lynch.2. Group48SPF male SD rats which wight from200-220g were randomly divided into control group, model group, Sanhuangyinchi decoction(SHYCD) high, medium and low-dose groups, Bicyclol group, each group of8.3. AHF model-makingAll rats were fed in SPF Barrier1week, then were fed drugs before being killed for5days. All rats were fasting but not water deprivation for12h before being killed. Then the rats in normal group were injected with normal saline while that in model group, SHYCD high, medium and low-dose group, Bicyclol group were injected with LPS(10μg/kg). After half an hour, the rats in model group, SHYCD high, medium and low-dose groups, Bicyclol group were injected with D-GalN(700mg/kg), then were Continue to feed drugs for treatmentl and provided a normal diet. All the rats were anaesthetized with sodium pentobarbital(400mg/kg), blood samples were collected by abdominal aorta and the serum were saved at-80℃after centrifuging.Livers were take out, part of them were soaked in4%paraformaldehyde, some were saved at-80℃.4. Indexs of observation and detectionThe serum level of ALT, AST, TBIL levels of rats in each groups were detected by automatic biochemical detector while the PT, INR and FIB were detected by automatic blood coagulation detector; the liver pathological morphology were observed by Nikon Eoipse Ti-s inverted microscope; the content of SOD and MDA in slurry of liver tissus were detceted by total SOD and MDA inmanufacturing detection kits; the expression of caspase3in livers were detected by Immunohistochemistry while the relative expression level of caspase3in livers were detected by Western blot.5. Statistical analysisThe data were statistically analyzed by using SPSS13.0software, One-Way ANOVA is applied to the compare of mean among several samples if multiple comparision among groups is in keeping with homogeneity of variance, if not, Dunnett will be applied. The value of P<0.05would mean significant difference.Result:1. Effect of SHYCD on AST,ALT and TBIL of AHF in ratsCompared with model group, SHYCD high, medium and low-dose groups and Bicyclol group both could reduce the content of ALT, AST, TBIL(P<0.01), SHYCD high-dose group was similar to Bicyclol group (P>0.05) while SHYCD medium and low-dose group were worse than Bicyclol group(.P<0.05).2. Effect of SHYCD on PT INR and FIB of AHF in ratsCompared with model group, SHYCD high,medium and low-dose groups and Bicyclol group both could shorten the time of PT and INR and both could increase the content of FIB(P<0.01), SHYCD high-dose group was similar to Bicyclol group (P>0.05) while SHYCD medium and low-dose group were worse than Bicyclol group(P<0.05).3. Histopathology of the liver in different groupsThe obversation of livers in each group by HE staining showed that:①in normal group, the structure of hepatic lobule structure is clear and the liver cells regular arranged;②in model group, the liver cells laminar necrosis, hepatic cords disorder, the structure of hepatic lobule structure is unclear; expansion,hyperemia and hemorrhage in hepatic sinus are obvious; inflammatory cell infiltration can be found within the hepatic lobule and collect abbacy; hydropic degeneration in the remaining liver cells;③the liver cells in test groups were improved, swelling alleviated, necrosis reduce, SHYCD high-dose group was similar to Bicyclol group while SHYCD medium and low-dose group were worse than Bicyclol group.4. Effect of SHYCD on SOD and MDA of AHF in ratsCompared with model group, SHYCD high, medium and low-dose groups and Bicyclol group both could increase the activity of SOD and reduce the the content of MDA(P<0.01), SHYCD high-dose group was similar to Bicyclol group (P>0.05) while SHYCD medium and low-dose group were worse than Bicyclol group(P<0.05).5. Effect of SHYCD on caspase3of AHF in ratsCompared with model group, SHYCD high,medium and low-dose groups and Bicyclol group both could reduce the expression level of caspase3(P<0.05), SHYCD high-dose group was similar to Bicyclol group(P>0.05) while SHYCD medium and low-dose group were worse than Bicyclol group(.P<0.05).6. Effect of SHYCD on cleaved caspase3of AHF in rats.Compared with model group, SHYCD high, medium and low-dose groups and Bicyclol group both could reduce the expression level of caspase3(P<0.01), SHYCD high-dose group was similar to Bicyclol group(P>0.05) while SHYCD medium and low-dose group were worse than Bicyclol group(P<0.05).Conclusion:1. SHYCD can improve the liver function and blood coagulation function.2.SHYCD can increase the activity of SOD, reduce the content of MDA, reduce the expression level of caspase3and inhibit the apoptosis of liver cells.3.SHYCD can prevent and treat the AHF and its possible mechanism is to inhibit caspase3stress and reduce liver cell apoptosis through the expression of antioxidant. |
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