It has been found that in recent years the incidence of liver diseases has been increasing greatly in Taiwan. Chronic liver diseases and liver cirrhosis are at the sixth among the top ten causes of death recently. Furthermore, liver cancer is the leading cause of cancer mortality. Liver resection and liver transplantation are the main therapy. Ischemia-reperfusion injury plays an important role during liver resection or liver transplantation. Ischemia-reperfusion (I/R) injury of the
liver may occur under many clinical conditions, such as hepatic trauma, hepatic transplantation, hypoperfusion shock or partial hepatectomy for liver tumors. Until now, the actual mechanisms remain unknown. There is evidenced that the sequence of hepatic I/R injury may cause severe liver injury. It was suggested that the increase of Kupffer cells activation induced reaction oxygen species involves the possible mechanisms. Activation of Kupffer cells results in production and release of
proinflammatory cytokins, including tumor necrosis factor α (TNF-α) and interleukin 1(IL-1β), chemokines, and neutrophil activation, lead to sinusoid endothelial cell death and hepatic cell damage. The period of hepatic ischemia associated with this technique and the resultant reperfusion can lead to liver injury and dysfunction, which is the main cause of death after hepatectomy. Thus, hepatic I/R injury has been actively investigated, and recently, protective strategies consisting of surgical
interventions, pharmacological agents, and gene therapy have been reported. Therefore, enhancing the safety of hepatic surgery and diminishing the significant rates of morbidity and mortality are the most important task in clinical therapy. In Taiwan, Antrodia camphorate(AC) is an exclusive fungus parasitic on the inner cavity of the endemic species Cinnamomum kanehirai Hayata and an important traditional Chinese medicinal fungus(Basidiomycetes) for the treatment of human disease such as
food and drug intoxication, abdominal pain, hypertension and liver cancer. Recently, polysaccharides extracted from fruiting bodies and mycelial cultures of Antrodia camphorate are reported to provide several therapeutic benefits including anti-inflammation, antioxidation, vasorelaxation and anti-hepatitis B virus activities, but the underlying molecular mechanisms are obscure. Our study had four groups which included ischemia reperfusion group、0.2 g/kg AC + I/R group、AC sham group and
normal control group. The I/R rats are treated with 0.2 g/kg Antrodia camphorate (AC) 30 minute before ischemia (IR, ischemia 30min + 0.2 g/kg A.C.), than reperfusion 6h or reperfusion 7 day. Rat serums which were assayed TNF-αquantity by ELISA were extrated before ischemia、after reperfusion 2h and 6h. The serums which were assayed IL-6 and IL-10 quantity by ELISA were extrated before ischemia、after reperfusion 1h, 2h, 3h, 4h, 5h and 6h. After reperfusion 6h the serums were also assayed
glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) quantity. Liver tissue【left lateral lobe(ischemia liver) and right lateral lobe(nonischemia liver】will be taken for measuring lipid peroxidation(MDA assay).liver tissue will also be taken for paraffin-embedded tissue sections which were respectively stained by H&E or TUNEL, and observed histopathology and liver cell apoptosis. In these results, the ischemia reperfusion animal modal was successful，when MBP
decreased at ischemia stage and increased gradually at reperfusion stage. Furthermore the liver was black and necrosis during reperfusion 6h. Pretreated Antrodia camphorate, we detected that rat GPT concentration was improved at reperfusion 6h stage, contrasted with IR control (P＜0.05), but GOT concentration was decreased slighter in pretreated Antrodia camphorate. The paraffin-emmbedded tissue sections which were stained by H&E showed that AC could diminish hepatocyte vacuolar degeneration,
vascular congestion and less severe hepatic necrosis in liver grafts at reperfusion 6 hours. The physiology monitory result showed : 1. After ischemia injury, AC could avoid high body temperature. 2. At ischemia stage, AC could maintain heart rate stability. 3. Contrasted with IR control, AC could maintain blood pressure stability at reperfusion 3~6 h. Under other hand, AC could reduce the TNF-α、IL-6 quantity of liver ischemia reperfusion injury rat serum, and suppress inflammation reaction. In
AC therapy group, because inflammation reaction were be suppressed, IL-10 quantity range on the rise were be down regulating in serum. When we assayed MDA level., which was decreased at Antrodia camphorate pretreatment, compared with IR control after reperfusion 6h. In the result of TUNEL stain, AC reduced severe hapatic cell DNA fragmentation and apoptosis in liver at reperfusion 6 hours. At reperfusion 7 day, we found AC which could increase the survival rate to 80％。 In this study, we had
established rat liver ischemia reperfusion modal. The results suggested, effect of Antrodia camphorate precondition to protect was good, which could reduce liver ischemia reperfusion injury. In the future, we will investigate the mechanism of protection with Antrodia camphorate, and add other medicine or therapy method which can increase the curative effect of liver ischemia reperfusion injury.
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