| The liver is the most important organs of alcohol metabolism,90%to95%ethanol oxidative metabolism in it. Long-term drinking a variety of alcoholic beverages can cause alcoholic liver disease. With the increasing number’of our drinking, alcohol abuse and alcohol dependence has become China’s increasingly serious public health problem. Alcohol on the liver injury involves multiple mechanisms, oxidative damage is recognized as one of the reasons. It is the main performance that the destruction of the liver’s redox environment, high generation of reactive oxygen species(ROS), lipid metabolic disorder, as well as biological macromolecules oxidative degeneration and the formation of adducts. All of these performance lead to the liver cell injury.Ginger has a long histery as medicine. Modern study found that ginger has anti-oxidation and reducing blood lipids of a variety of effects, conducive to the prevention of liver disease and cure. The gingerol has been proved to have various biological activities such as anti-inflammatory, oxidation resistance, anti-mutagenic, reducing blood lipids and anti-atherosclerosis, anti-tumor and etc., which is spicy ingredient of the ginger. This research want to find the best antioxidative active region of ethanolic extract from ginger, and then, find the protection of the best antioxidative active region to alcoholic liver injury.Objective:This reseach to gingerol as an indicator, using macroporous resin to isolate the ginger ethanolic extract further, select the the best antioxidative active region, and this research plans to confirm the antioxidation of gingerol against ethanol-induced hepatocyte damage.Methods:1. Separation of ginger ethanolic extract:Choose AB-8macroporous resin as the stationary phase and different concentrations of ethanol as eluent to separate ginger ethanolic extract. We get water region,25%ethanol region,50%ethanol region,75%ethanol region,95%ethanol region.2. The test of gingerol in crude extract and separating product:The standard curve was drown according to the vanillin as the standard by UV-visible spectrophotometry. Gingerol in crude extract and separating product were tested. 3. The test of gingerol in crude extract and separating product:For comparing the inhibition rate to liver homogenate MDA formation of crude extract and separating products, mouse liver homogenate was induced lipid peroxidation occurs, and compared with vitamin E; Testing the similar SOD activity of crude extract and separating product by SOD kit. On the basis of the content of gingerol and antioxidant activity, we got the best antioxidative active region of crude extract.4. Effect for antioxidant system of hepatocyte and Protection of gingerol:①Establish the hepatocyte oxidative damage model:In this study, we use diffent concentration ethanol to stimulate hepatocyte, and assemble cell at different time. Then we determine the activity of LDH, AST which released from hepatocyte, and determine the cellular MDA, SOD, GSH activity in order to observe the time-dose-response effects and establish the hepatocyte oxidative damage model.②Protection of gingerol for ethanol-induced hepatocyte damage:According to the oxidative injury model parameters, we use gingerol to pretreat hepatocyte before ethanol affect, and observe the time-dose-response defend effects in order to confirm gingerol can defend against ethanol-induced hepatocyte damage.Result:The gingerol content (6.4%)in crude extract was determined, while water region1.3%,25%ethanol region11.5%,50%ethanol region34.7%,75%ethanol region32.3%, and95%ethanol region28.5%; About MDA inhibition rates and similar SOD activity,50%ethanol region had the best effect. The conclusion was that50%ethanol region is the best effective part of antioxidant activity.It was positive correlation between ethanol stimulation dose and time and degree of damage. The vigor of hepatocyte degrade and degree of lipid peroxidation augment as well as the antioxidase degrade along with the increase of ethanol stimulate dose and time, it was significant difference at100mmol/L or8h comparing with normal hepatocyte.The best antioxidative active region(gingerol) of ginger ethanolic extract can protect hepatocyte against ethanol-induced damage. Pretreatment of gingerol can reduce GSH consume and inactivity of antioxidase and lipid peroxidation, which were provoked by ethanol. At the other hand, Gingerol can increase the cell metabolic activity and balance the antioxidative system to protect hepatocyte against oxidative damage. |
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