| Objective:To explore the protective effect of Mongolian medicine Gurigumu-13 on non-alcoholic fatty liver disease(NAFLD)and its possible mechanism.Methods:(1)40 Wistar rats were randomly divided into blank control group,model group,Gurigumu-13 high concentration group(1.24 g/kg d),Gurigumu-13 medium concentration group(0.62 g/kg d),Gurigumu-13 low concentration group(0.31 g/kg d),a total of 5 groups,8 animals in each group,the blank control group was fed with ordinary diet for 8 weeks,and the model group and the drug group were fed with methionine choline Lack of feed(Methionine-and Choline-Deficent Diet,MCD)for 4 weeks,at the 5th week,2 animals in the model group were sacrificed for liver sampling,and the appearance changes were observed and histopathological sections were stained.After confirming that the modeling was successful,the administration group was given high concentration,medium concentration and low concentration by gavage,and the blank group and model group were given PBS by gavage.The body weight of rats was recorded every 2 weeks.At the ninth week,the animals were sacrificed,and the samples were collected,and the liver weight and liver index were determined.HE staining was performed on the liver,and serum liver function indexes and oxidative stress indexes of liver tissue were detected.The mRNA and protein expression levels of NF-κB and Nrf2 were detected at the same time;(2)Preparation of drug-containing serum:Wistar rats were randomly divided into blank group,high-concentration group,medium-concentration group,and low-concentration group.Blood was collected after 7 days to prepare drug-containing serum;(3)HepG2 cells were treated with different concentrations of palmitic acid,and oil red O staining,TG content,and Western blot experiments were used to screen the modeling conditions;after successful modeling,different concentrations of drug-containing cells were used.Serum acted on it for 24 hours,and the liver function indexes,Oxidative Stress Indicators,and the mRNA and protein expression levels of NF-κB and Nrf2 were detected.Results:(1)Wistar rats were fed with MCD diet for 4 weeks,and their body weight increased,and the appearance of liver became fatty.Pathological staining showed that hepatocytes contained lipid droplets of different sizes,and the nucleus was squeezed to one side.The body weight and liver index of the rats in the model group were higher than those in the blank control group,indicating that the modeling was successful;(2)After 4 weeks of intragastric administration,with Compared with the model group,HE staining showed fewer lipid droplets,decreased TC,TG,LDL-C,ALT and AST indexes,increased HDL-C,and decreased oxidative stress indexes GSH-PX and MDA in the high and medium concentration administration groups.SOD and GSH increased;qPCR results showed that,compared with the model group,the expression of NF-κB mRNA in the high and medium concentration drug groups decreased,and the expression of Nrf2 increased,while the Western-blot results showed that the high and medium concentration drug groups NF-κB protein expression decreased,and the expression of Nrf2 increased(P<0.05);(3)After treatment of HepG2 cells with palmitic acid at a concentration of 0.25 μmol/L for 48 h,lipid droplets were accumulated in the cells,which proved that the in vitro cell modeling was successful.At the same time,the content of TG increased and The expression of NF-κB protein increased,while the expression of Nrf2 protein decreased(P<0.05);(4)After palmitic acid-induced HepG2 cells were treated with medicated serum,lipid droplets in the high and medium concentration medicated serum groups decreased,and the results of related indicators were the same as those of animals.The experimental results were consistent.The qPCR results showed that the expression of NF-κB mRNA decreased and the expression of Nrf2 increased in the high and medium concentration drug-containing serum groups.(P<0.05).Conclusion:Mongolian medicine Gurigumu-13 has a certain protective effect on NAFLD,and its possible mechanism is to promote the expression of antioxidant gene Nrf2 and inhibit the expression of NF-κB. |
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