| Objective: Zhige Oral Liquid is a new kind of solution wine to protect liver preparations,created by the professor sun of traditional Chinese medicine according to clinical experience for many years,this experiment using Zhige Oral Liquid intervention in the rat model with alcoholic liver disease,from Adenosine Monophosphate-activated Protein Kinase/sterol regulatory element binding transcription protein-1/Lipin-1(AMPK/SREBP-1/Lipin-1)pathway to explore the effect of Zhige oral liquid on glycolipid metabolism of rats with alcoholic liver disease.Methods: 1.the experimental group and building the model of alcoholic liver disease: 72 male SD rats clean,weight 200~230g,adaptive feeding 1 week first,and then were randomly divided into six groups: normal group,alcoholic liver disease model group(model group),Zhige Oral Liquid of high,medium and low dose group(here in after referred to as the high dose group,middle dose group and low dose group)and JieJiuLing oral liquid in the control group therapy(control group),each group of 12.Normal group rats lavage distilled water 1.0mL/100 g one day,according to the equivalent dose groups rats(1.0mL/100g/d)for luzhou is tempting lavage(alcohol content of 52%),alcohol lavage after high,medium and low dose group were given 1.0mL/100g/d,0.5mL/100g/d and 0.25mL/100g/d Zhige Oral Liquid lavage,while control group according to 0.5mL/100g/d lavage JieJiuLing oral liquid,for building and for lavaging 12 weeks.2.Materials: rats in each group were anesthetized by intraperitoneal injection of 10% chloral hydrate 0.3ml/100 g according to their body weight.After satisfactory anesthesia,blood was collected from abdominal aorta for centrifugation and stored in a refrigerator at-80?.The liver was cut into two parts and fixed with 10% formalin and 4% formaldehyde solution,respectively.3.Index detection: serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),triglyceride(TG),total cholesterol(TC),fasting blood glucose(FBG)and other index contents were detected by Enzyme standard instrument,and insulin concentration(INS)was detected by ELISA.Insulin resistance(IR)was calculated according to HOMA-IR mode [IR=(FBG*INS)/22.5],and insulin sensitivity index(ISI)was calculated according to HOMA-ISI mode [ISI=1/(FBG*INS)].HE staining and oil red O staining were used to detect the degree of liver cell damage and fat content.Immunofluorescence and immunohistochemistry were used to detect the expression levels of proteins involved in the AMPK/SREBP-1/Lipin1 pathway.Results: comparison of liver function indexes(ALT,AST): compared with the normal group,the ALT and AST levels of rats in the model group were increased(P<0.01),and the ALT and AST levels in the treatment group were decreased(P<0.01).The medium dose group was similar to the control group.Lipid metabolism related index(TC,TG,HE staining,oil red O staining)comparison: compared with normal group,model group rats TC and TG were higher(P<0.01),the treatment group rats blood lipid levels were falling compared with model group,which is still in high dose group had the greatest reduction(P<0.01),followed by the dose group and the control group;HE staining of liver tissue showed that: in the normal group,liver cells were of uniform size,clear boundaries,and normal morphology,with no fat vacuoles in them.In the model group,the liver tissue structure was disordered,the boundary was unclear,the nucleus was clearly shifted,and the distribution was uneven.A large number of fat vacuoles were observed in the whole liver tissue,and some small vacuoles were fused into large fat vacuoles.In the high-dose group,the cell structure was clear and close to that of the normal group.The performance of the medium dose group was about the same as that of the control group.There was little change between the low dose group and the model group.Oil red O staining showed that: normal group: hepatocytes were normal in size,nuclei were blue in color,cells were arranged in a regular and orderly manner,and no red lipid droplets were deposited in cells.In the model group,the liver cells were obviously swollen,and large red lipid droplets were observed around them.Compared with the model group,the red lipid droplet of liver cells in the high-dose,medium-dose and control groups of zhige oral liquid decreased,and the decrease was the most obvious in the high-dose group.The number of red lipid droplets in the liver cells of the medium dose group and the control group was similar,and the deposition of red lipid particles in the low dose group was significant,but still decreased compared with the model group.Comparison of glucose metabolism indexes(FBG,INS,IS,IR)in each group: Compared with normal group,model group rats FBG,INS content increased(P<0.01),liver on insulin sensitivity(IS)reduced(P<0.01),insulin resistance(IR)significantly(P<0.01),compared with model group,the rest of the serum levels of FBG and the INS all have varying degrees of decline,which the insulin sensitivity high dose group increased(P<0.05),insulin resistance improvement(P<0.01),almost close to the normal group,in general,the treatment group improved indicators of high dose group>the dose = control group>low dose group.Comparison of the expression levels of AMPK/ SREBP-1/Lipin-1 in liver tissues: compared with the normal group,the expression levels of AMPK in the model group were significantly inhibited,and the expression levels of SREBP-1,lipin-1 and FASN in the downstream lipid proteins were increased(P<0.01).Compared with the model group,the liver AMPK protein was activated in each treatment group,and the expression of its downstream lipid protein was inhibited.The difference was the most obvious in the high-dose group(P<0.01),followed by the medium-dose group and the control group(P<0.05),and the improvement was not significant in the low-dose group(P>0.05).The same result can be obtained by optical density analysis of immunohistochemistry..Conclusion: 1,Zhige Oral Liquid can reduce the rats with alcoholic liver disease the liver damage,reduce blood fat levels,reduce liver lipid deposition and fat vacuoles,and can reduce the alcoholic liver disease in rats blood glucose and blood insulin levels,increased insulin sensitivity of the liver,improve insulin resistance,thus regulate glucolipid metabolic balance;2,its mechanism may be related to AMPK/ SREBP-1/Lipin-1 glucolipid metabolic pathways,Zhige Oral Liquid can be expressed through the activation of AMPK,cut its downstream lipid protein FASN,SREBP-1,Lipin-1 synthesis,improving the body condition,high cholesterol,high blood sugar,high insulin concentrations and glucolipid metabolism in the rat model of alcoholic liver disease,prevention and treatment of alcoholic liver disease development. |
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