| Objective: Non-alcoholic fatty liver(NAFLD)model rats were used as research objects,and the mechanism of non-alcoholic fatty liver disease was studied based on the intestinal flora-based Paeonal(Pae).Method: In this study,a non-alcoholic fatty liver rat model was replicated by intragastric administration with a high-fat emulsion,and the activity and pathological condition of the rats after drug intervention were observed.SPF grade SD rats were randomly divided into normal group and model group.In nine weeks,the model group was given high-fat emulsion but the normal group was given normal saline.After successful modeling,the model group rats were further randomly divided into model control group,positive drug group,and Paeonal(Pae)groups with high,middle and low dosages respectively.The activity of rat models and medication was observed by gavaging the high-fat emulsion in the morning while treating Paeonol in the afternoon all in 4 weeks.After 4 weeks of treatment,all the rats were sacrificed and the pathological changes of liver tissue were observed under light microscope.In rat liver tissue,Enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of inflammatory factors IL-1beta,IL-2,IL-6,IL-8,TNF-alpha and NF-kappa B and IL-2,IL-6,TNF-α and NF-κB mRNA expression was find with qRT-PCR method;the content of free fatty acid(FFA)in serum and liver tissues of rats was detected by ELISA;the expression of NF-κB in liver tissue was detected by immunohistochemical DAB staining;the superoxide dismutase in liver tissue of rats was detected by WST-8 method.;the content of SOD was detected by Western blot;the expression of IKBα and NF-κBp65 protein was detected by Western blot;the composition of intestinal microbiota was analyzed by sequencing technique.Results: This article is to induce the formation of nonalcoholic fatty liver in rats by intragastric administration of high-fat emulsion.Compared with the normal group,the rats in the model group had loss of appetite,dull hair,weakened activity,and the stoolwas soft and smelly.After the drug intervention,the hair and activity of the positive drug group and the paeonol high-dose group were significantly improved.Under the light microscope,the liver cells of normal rats were found to have no steatosis.The hepatocytes of the model group showed significant fatty degeneration.Compared with the model group,there are obvious improvements in the treatment group with decreased hepatic steatosis as well as little lipid droplets though.The results of ELISA showed that the levels of inflammatory factors IL-1β,IL-2,IL-6,IL-8,TNF-α and NF-κB were statistically different between the groups;gene expression analysis showed that each drug was used.The expression of IL-2,IL-6,TNF-α and NF-κB mRNA in the liver tissues of the rats was significantly down-regulated.The expression of NF-κB in the model group was positive by immunohistochemical staining.The content of free fatty acid(FFA)in the serum and liver tissues of rats was determined by ELISA.The content of FFA in the serum and liver of the model group was normal.The group was significantly elevated.The content of superoxide dismutase(SOD)in the liver of rats was detected by WST-8 method.It can be seen that the content of SOD in the model group was significantly decreased compared with the normal group.The WB method was used to detect IKBα and NF-κBp65.Protein content,the content of IKBα in the model group decreased significantly,and the content of NF-κBp65 increased.Rat feces were taken,and the composition of intestinal microbiota was analyzed by sequencing technology.In the Bar chart of community analysis,the content of Firmicutes was higher at gate level,and the difference of dominant genera among CG,MG,TG and PG groups was analyzed at the level of significant difference test genera.The results showed that unclassified-f-Lachnospiraceae,unclassified-p-Bacteroidales,Lachnospiraceae-ND3007-group had significant or extreme differences among the four groups.The content of Paeonol in the model group was higher than that in the normal group,and it decreased after paeonol treatment.Conclusion: In this study,the production of nonalcoholic fatty liver in rats was induced by intragastric administration of a high-fat emulsion.Paeonal(Pae),bysequencing the intestinal microflora of feces in non-alcoholic fatty liver rats,obtained the content of Firmicutes in the model group of the four groups of midgut microbial flora at the gate level of the community histogram.Higher.Compared with the normal group,the relative abundance of the genus Trichophyton was higher in the model group,and decreased after treatment with paeonol;paeonol inhibited liver inflammation in rats with nonalcoholic fatty liver The release of factors significantly improves the levels of inflammatory factors,free fatty acids,superoxide dismutase,etc.in rats,and achieves the purpose of treating liver tissue damage. |
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