| A Nonalcoholic fatty liver disease(NAFLD)refers to non-alcohol and other specific factors that caused liver damage,Closely related to heredity,environment and metabolism,including nonalcoholic fatty liver disease,nonalcoholic steatohepatitis(NASH),nonalcoholic fatty liver hepatic fibrosis and cirrhosis.In recent years,with the change of people's living habits and dietary structure,the incidence of NAFLD has been rising and younger,which has become one of the global public health problems in the twenty-first Century.With the change of people's living habits and dietary structure,the incidence of NAFLD is rising and appearing more in younger people.The overall incidence of non-alcoholic fatty liver disease has reached 20%-30%and 12%-24%in Western countries and in Asia respectively.The incidence has apparently increased in the past fifteen years,but its pathogenesis has not been fully elucidated.In recent years,it has been discovered that the formation of NAFLD and peroxisome proliferator activated receptor in the process(Peroxisome proliferator-activated receptor alpha,PPAR alpha,uncoupling protein 2(Uncoupling)protein,UCP-2)and peroxisome proliferator activated receptor activation on-1 alpha-Y auxiliary(Peroxisome proliferator-activated receptor gamma coactivator-1 alpha,PGC-1 alpha)plays an important role.Its PPAR alpha and PGC-1 alpha signaling pathways are directly involved in the metabolism of brown and white fat,contributing to fat and energy metabolism,thereby reducing hepatic lipid deposition.The whole world is also a lack of drug efficacy,safety and stability in the treatment of NAFLD,so the study of national drug intervention for PPAR alpha and PGC-1 alpha signaling pathway,promote fat metabolism,to achieve the purpose of drug treatment of NAFLD is innovative.Nonalcoholic fatty liver disease is diagnosed as "Tonglaga undigested disease" category of Mongolian medicine."Tonglaga undigested disease" is mainly due to excessive eating greasy,cool,not easy to digest food and other incentives caused by voicing biochemical dysfunction caused by accumulation of fat in the liver disease characterized by.Bright four salt flavor soup is the preferred treatment of "Tonglaga undigested disease",and in fat,have a certain effect to promote fat metabolism,especially the monarch drug composition of Piper longum and dried ginger in both glucose metabolism and antioxidant activity of fat metabolism,strong regulation.This experiment illustrates bright four salt flavor soup to promote fat metabolism,molecular mechanism of inhibition of fat deposition in the liver,for clinical rather widely used bright four salt flavor soup and from Mongolian drug development to provide a scientific basis for the treatment of NAFLD.Purpose:To observe the effect of bright four salt flavor Decoction on nonalcoholic fatty liver model in mice body weight,liver index,liver steatosis,liver function and influence on the expression of PPARa and UCP-2 gene in mRNA adipocytes and PGC-la,the efficacy of the treatment of verification of bright four salt flavor Decoction on nonalcoholic fatty liver disease,to explore the liver the protective effect and mechanism.Method:1.80 healthy male C57BL/6n mice were randomly divided into two groups:blank control group(15 mice)and high-fat model group(65 mice).The blank control group was given normal diet,and the high-fat model group was fed with high fat diet(Research,Diets,D12492;60,kcal%,Fat).After 12 weeks,3 mice were randomly selected from the blank control group and the high-fat model group.The liver morphology was observed and the level of fat deposition in the liver was confirmed.The replication degree of NAFLD model in C57BL/6n mice was evaluated.When the model was established successfully,the hyperlipidemia model mice were randomly divided into model group,tiopronin group(positive control group),bright four salt flavor Decoction group(observation group)and other three groups were continued to be fed with high fat diet and blank control group continued to be fed with normal diet.From the beginning of the thirteenth week,the blank control group and the model control group were given distilled water 10ml/Kg/d intragastric administration,the positive control group was given tiopronin 56mg/Kg/d intragastric administration,the observation group was given light salt four flavored soup 0.45g/Kg/d gavage.After continuous intragastric administration for 4 weeks,the serum and liver tissue samples were taken after anesthesia.Weighing the body weight and liver of mice,liver index,automatic biochemical detection of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST),r-glutamyltranspeptidase(gamma-GT),total cholesterol(TC),triglyceride(TG)content.HE staining and Oil red staining were used to evaluate the pathological changes of liver tissue in each group.2.3T3-L1 preadipocytes differentiated into mature adipocytes,and they were randomly divided into five groups:blank control group,PBS group(positive control group),Bright four salt flavor decoction,high,medium and low dose group(observation group).Do not add any drug control group,PBS group added 10%PBS buffer,bright four salt flavor soup is low,medium and high dose group were added with low dose(5mg/ml),middle dose(15mg/ml)and high dose(20mg/ml)bright four salt flavor soup,inside the bottle under the same conditions respectively.48 hours after culture,extracting total RNA and protein in fat cells,the detection of PGC-la,PPARa and UCP-2 implementation method of RT-PCR gene expression levels of mRNA expression by Western Blot detection of PGC-1 alpha and UCP-2 protein.All the values were expressed by meanąstandard error.The t test was performed by SPSS 19.00 software,When P<0.05,there were significant differences between the observation group and the control group.Result:Experiment 11.Oil red staining showed that compared with the blank control group,the staining degree of liver Oil and red in the model group was distinct,the staining area of liver was slightly weakened and the area became smaller in tiopronin group.The staining degree of Bright four salt flavor Decoction group was obviously lower than that of tiopronin group and model group,and the area of dyeing was small.2.compared with the blank control group,the liver index increased significantly in model group,(P<0.05);compared with the model group,bright four salt flavor Decoction group and tiopronin group liver coefficient decreased significantly and there was significant difference(P<0.05);compared with the blank control group,model group serum in ALT,AST increased significantly and there was significant difference(P<0.05);compared with the model group,tiopronin group,serum ALT,AST significantly decreased and there was significant difference(P<0.05);but compared with the model group,bright four salt flavor soup group serum ALT,AST significant differences(P>0.05);there was no significant difference between the groups in serum gamma-GT(P>0.05);compared with the control group,the model group was TC,TG was significantly increased and there were significant difference(P<0.05);compared with the model group,bright four salt flavor soup in serum The levels of TC and TG were obviously decreased(P<0.05),but there was no significant difference in serum TC and TG between the tiopronin group and the model group(P>0.05).Experiment 21.RT-PCR showed that:compared with the control group,PBS group of fat cells PGC-la gene expression of mRNA increased significantly and there was significant difference(P<0.05);compared with the PBS group,bright four salt flavor soup group,high dose group of fat cells PGC-1a gene expression of mRNA was significantly higher(and there is a significant difference P<0.05);compared with the control group,the expression of PBS PPARa gene of the mRNA group of fat cells had no significant difference(P>0.05);compared with PBS buffer group,bright four salt flavor soup group,high dose group of fat cells PPARa gene expression of mRNA increased significantly and there was significant difference(P<0.05);compared with PBS buffer group,the expression of bright four salt flavor soup fat cells in each group UCP-2 gene mRNA had no significant difference(P>0.05),but with the increase of bright four salt flavor soup dosage decreased.2.Western Blot assay results showed that:compared with the control group,PBS buffer group PGC-la protein expression increased significantly(P>0.05);compared with PBS buffer group,the expression of bright four salt flavor soup high dose group of PGC-1a protein increased significantly(P<0.05);compared with the PBS group,bright four salt flavor soup group UCP-2 protein expression increased with the bright four salt flavor soup dosage decreased trend.Conclusion:1.Bright four salt flavor Decoction can efifectively reduce the liver wet weight and liver index of non-alcoholic fatty liver model mice,and reduce the fat deposition in liver tissue.2.Bright four salt flavor decoction has obvious lipid-lowering and inhibit fat deposition in the liver of nonalcoholic fatty liver model in mice,and lipid-lowering effect is better than Tiopronin Enteric-coated Tablets.3.Bright four salt flavor decoction can activate the PGC-la/PPARa signaling pathway and correct the disorder of hepatic lipid metabolism.4.Bright four salt flavor Decoction mainly through the promotion of fat cell PGC-la and its receptor PPARa synthesis,up regulation of PGC-1a protein expression,thereby promoting energy and fat metabolism,prevent cell fat deposition. |
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