Studies On Protection Effect And Mechanism Of Ruwe In Injury Of The Mice Liver

Objective:Studies on the protective effect and mechanisms of Rhaponticum uniflorum water extract (RUWE) in injury of the Mice liver by CCl4.Methods:1、 Establish CCl4induced liver injury model, respectively, with different CCl4concentration and different time stimulating the mice to study the impact of CCl4on the mice. Kit to detect the activity of mouse serum alanin aminotransferase (ALT) and aspartate aminotransferase(AST) and get the best concentration and stimulation time of CCl4induced liver injury.2、acute liver injury:the mice were randomly divided into normal control group、model group、low、mediurn、high medication group(50、1O0、200mg/kg)、Silymarin group(100mg/kg); normal control group and Silymarin group fed with20mg/kg saline daily, Gavage for7days after the last administration8h,in addition to the normal group,other groups intraperitoneal injection of0.2ml of CCl4soybean oil solution32mg/kg,Fasting but water after16h to obtain blood and liver.subacute liver injury:grouping as same as the acute liver injury, Gavage for24days after the administration8h,in addition to the normal group,others groups intraperitoneal injection of CCI4soybean oil solution32mg/kg every three days, the last administration8h,Fasting but water after16h to obtain blood and liver.3. Kit to detect the activity of mouse serum alanine aminotransferase (ALT), aspartate aminotransferase(AST), superoxide dismutase(SOD)、hydrogen peroxide enzyme(CAT)、glutathione-s-transferase(GST) and glutathione peroxidase(GSH-PX),We also use kit to determine malondialdehyde (MDA) and glutathione(GSH) in liver homogenates. We use protein immunobltting detect the protein expression of therapeutic targets, such as, COX-2, iNOS and HO-1in microsome, Also use Western blot to detect the expression extracellular singal-regulated protein kinase(ERK),c-jun amino-terminal kinase(JNK), p38mitogen activated protein kinase(p38), nuclear transcription factor-KB(NF-KB)(including the nucleus and the cytoplasm)and its phosphorylated form.Results:32mg/kg CCl4induced liver injury in mice by intraperitoneal injection16h,ALT and AST levels were significantly higher than normal group (P<0.05);2.Give different concentration of RUWE, compared to model group,the activity of ALT and AST levels of treatment group with increasing drug concentration progressively reduced (P<0.01).In liver homogenates. The content of MDA progressively lower(P<0.05), but the GSH conversely(P<0.01); the activity of GSH-PX progressively increased(P<0.05), the activity of CAT and SOD progressively increased inliver homogenate(P<0.01).3. Western blot showed that in model group,the expression level of HO-1,COX-2and iNOS increased, the expression level of HO-1,COX-2and iNOS in medication group significantly reduced-CCl4can significantly elevated protein expression level of NF-κB、 p38、JNK、ERK and its phosphorylated form,compared with model group, protein expression level of NF-κB、 JNK in medication group were decerased, and protein expression level of p-NF-κB、p-JNK、p-p38significantly reduced(P<0.01), the expression of ERK and p-ERK no significantly change.Conclusion:RUWE has a significant protective effect on CCl4induced liver injury in mice,and significant inhibition of CCl4in mice liver injury.Its role may be related to increase antioxidant capacity,inhibit protein activation of COX-2、iNOS、p38、JNK、 NF-κB.