The Research Of Effectiveness Of Acupuncture On Blood Perfusion Flow And Oxidative Stress Reaction On Mice Of Alcoholic Liver Injury

Objective:Basis on the established imaging method of laser speckle imaging technique for hepatic blood perfusion to observe the hepatic blood perfusion differences between normal mice and acute alcoholic hepatic injury model mice, and the effort of Electroacupuncture (EA) act on the hepatic blood perfusion of both normal mice and acute alcoholic hepatic injury model mice; Explore the effect on biochemical criterion of oxidative stress through electroacupuncturing acute alcoholic hepatic injury model mice.Methonds:There are three parts in this paper. Part1. Preparation the model of acute alcoholic liver injury and observation of the hepatic tissue morphology.16mice are randomly assigned to normal control group and acute alcoholic liver injury model control group (model control group),8mice in each group. First is the preparation of acute alcoholic liver injury model. Each one of model control group is given15ml/(kg. bw) of50%ethanol by intragastric infusion, then they are absoluted diet but not water12h. Once that is done,8mices of each group are all put to death, then take off their heptic tissue immediately to do biochemical test of both MDA and SOD in order to validatie successful making of acute alcoholic liver injury model. Second is to observe the change of heptic tissue morphology through the heptic tissue HE staining slice of acute alcoholic liver injury model mice. Part2. Displaying the effect of acupuncture act on the heptic blood perfution of mice through laser doppler blood perfusion imaging technology. Put20normal mice and20model mice (According to the part one) divide into normal control group, normal acusector group (normal EA group), model control group and model EA group. First of all, On the basis of established laser speckle blood perfusion imaging display methods of mice liver, hepatic blood perfusion of10mice in both normalcontrol group and model control group are detected30min by laser speckle instrument for observeing the distribution and variation of heptic blood perfusion. Secondly, hepatic blood perfusion of10mice in both normal EA group and model EA group are detected30min by laser speckle instrument, meanwhile received stimulation by electric acupuncturing (EA)"Taichong"(LR3) and "Zusanli"(ST36)30min for observeing the effect of acupuncturing action on heptic blood perfution. Then extract and analyze data through the related software of laser speckle instrument to observe differences of heptic blood perfution of normal control group, model control group, normal EA group and model EA group mice. Part3. The effect of pathological indicators of oxidative stress reaction of acute alcoholic liver injury model mice induced by EA. Divide8normal mice and8model mice (According to the part one) into normal EA group and model EA group. All mice in each group are put to death and take off their hepatic tissue immediately to do biochemical test of both MDA、SOD and HE staining slice as part1. The first is to observe the HE staining slice through light microscope, then MDA and SOD in hepatic tissue of above-mentioned four group (normal control group, model control group, normal EA group, model EA group) are compared respectively to study the regulating action for pathological indicators of oxidative stress reaction by EA. Results:Part11. The hepatic tissue slices with HE staining of model mice showed the hepatic tissue has pathological change significantly.2. The MDA level of model mice is higher significantly than normal mice, the SOD level of acute alcoholic liver injury model mice is lower remarkedly than normal mice.Part21. Manifestations of laser speckle hepatic blood perfusion image of model mice:The hepatic blood perfusion images of model control group mice displayed the hepatic blood perfusion is lower than normal control group mice. Hepatic blood perfusion of the perimeter zone of model control group is lower than porta hepatic and the central zone, during the30min of detection, the hepatic blood perfusion with small fluctuation; The hepatic blood perfusion images of normal control group mice displayed the hepatic blood perfusion is abundant, blood perfusion of the perimeter zone of these livers are lower than porta hepatic and the central zone. During the30min of detection, the hepatic blood perfusion with small fluctuation.2. Manifestations of laser speckle hepatic blood perfusion image of mode mice by EA:during the30min of EA, hepatic blood perfusion of both mode mice and normal mice could be increased gradually by EA.3. The quantitative analysis of hepatic blood perfusion:Hepatic blood perfusion of mode control group mice is lower remarkable than normal control group mice(P<0.05); Hepatic blood perfusion of model EA group mice get the highest while at EA25min point, it can increase by (9.23±12.79)%compared with0min point. Hepatic blood perfusion of model EA group only at25min point has significant difference compared with0min point (P<0.05). Each viewing time of hepatic blood perfusion of model EA group has significant difference compared with the same viewing time of model control group (P<0.05); Hepatic blood perfusion of normal EA group mice get the highest while at EA25min point, it can increase by (12.44±10.92)%compared with0min point. Except at5min point and10min point, each viewing time of hepatic blood perfusion of normal EA group has significant difference compared with0min point (P<0.05). Except at10min point, each viewing time of hepatic blood perfusion of normal EA group hasn’t significant difference compared with the same viewing time of normal control group(P>0.05).Part31. The hepatic tissue slices with HE staining of model EA group showed the pathological change has reduced compared with model control group.2. The quantitative analysis of MDA in hepatic tissue:The MDA level of model EA group is lower remarkedly than model control group (P<0.05); The MDA level of normal EA group is lower remarkedly than normal control group (P<0.05).3. The quantitative analysis of SOD in hepatic tissue:The SOD level of model EA group is higher sightly than model control group (P>0.05); The SOD level of normal EA group hasn’t remarked difference compared with normal control group (P>0.05).Conclusions:1. The hepatic tissue has pathological changes significantly, the MDA level increases significantly and the SOD level decreases remarkedly by giving15ml/(kg. bw) of50%ethanol by intragastric infusion and absoluting diet but not water12h to Kunming mice. 2. Hepatic blood perfusion and the effects of EA on hepatic blood perfusion can be recorded accurately and displayed visually by laser speckle blood perfusion imaging.3. Hepatic blood perfusion of both model mice and normal mice can be increased induced by EA.4. The pathological change of the hepatic tissue in model mice can be repaired to some extent by EA.5. The MDA level of model mice can decrease through EA, and the SOD level can increase; The MDA level of normal mice can decrease through EA, but the SOD level variate small.