Protective Effect On Alcoholic Liver Injury Of Polyphenols From Punica Granatum L.

Tunica granatum L. is also called pomegranate etc., Pomegranate belongs to the genus Punica from the family Punicaceae, which is a very rich plant resources and widely distributed in China. Modern research has showed that:the effective components of pomegranate leaves contained polyphenols, polysaccharides, flavonoids etc., given pomegranate leaves have antioxidant, antibacterial, antiviral, convergence anti diarrhea, reduce blood fat and other physiological effects. Therefore, the purpose of this experiment was to extract polyphenols from pomegranate leaves, and the protective effect of polyphenol from pomegranate leaves on alcoholic liver injury was explored in the view of antioxidant, biochemistry, liver tissue pathology and ultrastructural observation.1. Polyphenols from Pomegranate leaves was extracted and the compent of Polyphenols was verified. The best conditions of extraction (the solid-liquid ratio,, extraction temperature, extraction time, extraction of pH)were studied by single factor test and response surface method, then CPPL were isolated, purified by macroporous resin, the pomegranate leaf polyphenols 1 (PPPL1) was got, PPPL1 were further isolated by polyamide column chromatography, the higher purity of pomegranate leaf polyphenols 2 (PPPL2) was got. The results of the optimum technological conditions of CPPL shpwed that the solid-liquid ratio was 20:1, extraction temperature was 60℃, the extraction time was 1.5h, the content of CPPL reached to 265.35mg/g; The purification results of CPPL showed that the most suitable macroporous resin purification of CPPL was AB-8, the static adsorption conditions:the concentration of sample was 2.5mg/mL, pH was 4, the sample rate was 2mL/min, desorption conditions were:60% alcohol, the purity of PPPL1 reached 62.44%, the purity of PPPL2 reached 92.83%; high performance liquid chromatography technology verified that PPPL2 contained gallic acid and ellagic acid and other polyphenols.2. The improvement of pomegranate leaf polyphenol on acute alcoholic hepatic injury in mice was investigated from the perspective of antioxidant, and scavenging free radicals of pomegranate leaf polyphenols and the ability of antioxidation in vivo were studied. First of all, the scavenging action of CPPL, PPPL1, PPPL2 on superoxide radical, hydroxyl radical and reducing power, nitrite were respectively determined. The results showed that: CPPL, PPPL1, PPPL2 had reducing capacity, better inhibition of nitrite and the ability of scavenging superoxide radical, hydroxyl radical, its antioxidant effect: PPPL2>PPPL1>CPPL. Then the improval effect of PPPL2 on acute alcoholic hepatic injury by the establishment of acute alcoholic liver injury in mice was investigated.60 mice were randomly divided into control group, alcoholic model group, the low, middle and high dose of polyphenols from pomegranate of leaves groups ((100> 200> 400) mg/ kg-bw)), positive control group (bifendate,150 mg/kg·bw), each group has 10 mice. The mice of control group and model groups were filled with deionized water (0.1 ml/10g·bw) daily, the mice were intragastric administration for 30 days. The mice of three different doses groups were filled with polyphenols from pomegranate leaves ((100、200、400) mg/kg-bw)). The positive control group mice were filled with bifendate (150 mg/kg·bw). The model of acute alcoholic liver injury was established on the 31st day,all mice were filled with 50% alcohol,except the control group, after 12 h, the mice in ocular blood, serum, liver were got. The results showed that catalase in liver and serum of model mice CAT activity, SOD activity, GSH-Px activity and GSH content was lower than that of control group, the MDA content increased significantly, while the different does of PPPL2 can increase in liver and serum of mice CAT, SOD, GSH-Px activity and GSH content, decrease the content of MDA, inhibite lipid peroxidation in varying degrees, some indexes of the high dose of PPPL2 even have reached the level of positive control group. Above results suggested that pomegranate leaf polyphenols not only has good antioxidant capacity in vitro, but also has a certain protective effects on alcoholic liver injury. The reason of protective effects maybe has a relationship with PPPL can enhance the body’s antioxidant activity, improve the body’s antioxidant capacity, improve low antioxidase activity in vivo due to alcohol.3. The regulating effect of PPPL2 on mice of TG ability, ALT ability, AST ability by the establishment of acute alcoholic hepatic injury in mice were investigated, and the improvement of PPPL2 on alcoholic liver injury in mice was studied. It can easily discover that the ability of ALT, AST in the model group mice liver and the ability of ALT, AST TG in serum levels were significantly increased, while the PPPL2 groups had a certain improving effect on alcoholic liver injury in mice, and the more dose the better effect, the ability of the liver ALT and serum TG, AST, ALT of high-dose PPPL2 had reached the level of positive control group; Pathological section indicated that liver cells were necrotic, inflammatory infiltration of liver cells were bad, liver cords disorders severe, while the mice of PPPL2 groups can gradually improve the liver inflammation, alcoholic liver fatty, hepatic cord disorders and other illnesses with the dose increased; The ultrastructure of hepatic tissue in model group mice showed that:liver cell of the mice appeared a number of lipid droplets, nuclear membrane shrank dramatically, the chromatin was in the abnormal distribution, mitochondria cristae blurred, rough endoplasmic reticulum was expanded seriously, ribosomal was off. It can be seen that PPPL2 has certain protective effects on alcoholic liver injury.