Effects And Mechanism Of Indole-3-carbinol And Sodium Ferulate Against Oxidative Damaged Liver In Rats

During the development of hepatic damage and diseases induced by various causes, rised oxidative stress and accumulation of lipid peroxidant are the common mechanism and pathological pathway. Aiming to the mechanism to conduct effective antioxidant investigation is one of main currents in the treatment of the hapatic diseases. Indole-3-carbinol (I3C), one of the glucosinolate compounds in cruciferous vegetables, can strengthen the capacity of antioxidative enzymes in various organs and cells. Sodium ferulate (SF), extracted from DangGui, containing phenolic hydroxyl has the antioxidative activity. This study has investigated the above two drugs' effects on damaged liver precision-cut slices and isolated active hepatic stellate cells (HSC) and related pharmacological mechanisms using technologies of liver precision-cut slices and cell culturing so that drugs in protection of damaged liver can be effectivly researched and selected. At the same time, this study has tried to establish the basement to smoothen the way of subsequently further exploring of HSC's active mechanism and accordingly therapeutic measurements.AIM To establish model of ethanol-induced hepatotoxicity in precision liver-cut slices of rat and to study the hepato-protection of DC and SF. To establish method of isolating HSC and model of isolated rat HSC auto-oxidative activation in vitro and investigate effects of DC and SF on cell proliferation .METHOD Ethanol 50 mmol ?L"1 was used to induce hepatotoxicity in precision liver-cut slices of rats. The release of glutamic pyruvic transaminase (GTP), glutamic oxaloacetic transaminase (GOT), lactate dehydrogenase (LDH) and glutathione S-transferase (GST) into culture medium, and aniline hydroxylase (ANH) and alcohol dehydrogenase (ADH) in cytosol of hepatocytes in the precision liver-cut were measured with different concentrations of DC and SF. The histopathological observation of liver slices was also done to evaluate the effect of DC. Use method of pronase E and collagenase IV perfusing and one step density centrifugation of nycodenze with a little modification according to references and conditions of laboratoryto isolate HSC. HSC was identified by optic and fluorescent microscope. Cell viability and yield was evaluated by trypan blue exclusion and purity of cells was assessed by the positive immunofluorescence staining for Desmin. Then to establish the auto-oxidative activation model of HSC and observe the effects of 13 C and SF with different concentrations on proliferation of activated HSC. RESULTS The releases of GTP, GOT, GST and LDH into the medium increased significantly when the slices were treated with 50 mmol ?L"1 ethanol for 4 h. DC at concentrations of 100 ~ 400 umol ?L"1 and SF at concentration of 463 ~ 1388 nmol ?L"1 effectively reduced the release of GTP, GOT, GST and LDH in the medium of ethanol-treated slices. Meanwhile, the activities of ANH and ADH in cytosol decreased to the normal levels. The histopathological changes of liver slices induced by ethanol were lessened by I3C treatment. Cell yields in each liver is 0.5- 1.2X 107/liver. The average viability by trypan blue exclusion was 94%. The grown cell purity for 24 hours tested by immunocytochemical phenotyping for desmin protein was 95%. I3C whose concentration were between 100 and 800 umol i/1 could significantly inhibit the proliferation of HSC in a dose-dependent manner. Concentrations of SF with 231 ~ 463 nmol ?L"1 could also inhibit the proliferation of HSC. CONCLUSION 13C and SF can markedly protect rat liver slices from ethanol-induced hepatotoxicity, which partly results from the change of ethanol metabolism in liver. The technique for isolation of HSC described here is successful and reliable. The auto-oxidative activated HSC model settled well. I3C and SF can significantly inhibit the proliferation of activated HSC.