Study On The Intervention And Tissue Differential Expression Of Aerobic Exercise And Black Fruit Wolfberry Flavone On Alcoholic Liver Injury Mice Based On NF-?B Pathway

Objective:In recent years,Alcoholic liver disease(ALD)has attracted much attention.At present,the treatment of alcoholic liver injury has not yet been a good solution,for this study of alcohol-induced liver injury in the occurrence and development mechanism,to explore effective treatment is the key to the clinical treatment of such patients.To investigate the effects of modeling and related stress factors in mice with alcoholic liver injury.To study the effect of aerobic exercise combined with black fruit wolfberry flavone on alcoholic liver injury.On this basis,the gene expression profiles of NF-?B pathway and its related factors in liver and blood tissues of mice were analyzed,and the effects of aerobic exercise and black fruit wolfberry flavone on alcoholic liver injury in mice and its related inflammatory mechanism were discussed.Methods:Fifty healthy Kunming mice,male and SPF grade,were used to establish the model of alcoholic liver injury by oral administration.The normal group was perfused with equal volume of distilled water,and the experimental mice were randomly divided into model after divided into model group,flavone group,aerobic exercise group and exercise+flavone group.Each intervention group began to conduct aerobic exercise and black fruit wolfberry flavone on the third day after the successful modeling,at a fixed daily time point.Among them,the aerobic exercise of mice was done by running on the platform and doing moderate aerobic exercise with zero slope;black fruit wolfberry flavone daily oral administration with a concentration of 200mg/kg,gavage 1 times.All interventions were 6 times a week for 6 weeks.During the intervention period,the mice were subjected to general behavioral observation.After the experiment,all the mice fasted overnight,weighed the anesthetized animals and killed,isolated the liver tissues and weighed them.The liver tissues of the mice were observed by microscopic morphology and histopathological score by HE staining.The expression of NF-?B,TNF-a,IL-6 cytokines in mouse liver was detected by immunohistochemical method,the expression of NF-?B,TNF-a,IL-6 and IL-10 in liver and blood tissues were detected by real-time fluorescence quantitative PCR,and the contents of alt and TBIL were detected by blood biochemistry.Results:(1)The results of serology:Compared with the model group,the content of ALT?AST and TBIL in the three groups was significantly lower than that in the model group(P<0.01),and the contents of the three components in the aerobic exercise group had a decreasing trend(P<0.05).(2)HE staining and histopathologic score showed that the pathological changes were mainly steatosis of liver cells.Compared with model group,the steatosis scores in each intervention group were statistically significant(P<0.01).The normal group of mice had complete liver cell structure.In model mice,the hepatic cord structure was disordered,the lobular structure was seriously destroyed,the cytoplasm was loose and the fat vacuoles were filled with different sizes,and accompanied by inflammatory cell infiltration.Aerobic exercise group,flavone group liver damage in mice improved to varying degrees,reduced inflammatory cells.The lobule structure of liver tissue of mice treated with exercise+flavone group was almost complete,the necrosis of liver cells and fat accumulation were alleviated,and the infiltration of inflammatory cells decreased rapidly(P<0.01).(3)The results of immunohistochemistry showed that the expression of NF-?B,TNF-a and IL-6 were the highest in the liver of the model group.Compared with the model mice,the aerobic exercise group,the flavone group and the exercise+flavone group had statistical significance(P<0.05,P<0.01).the aerobic exercise group,the black citrate flavonoid group and the exercise flavone group.(4)The results of Real-time PCR detection showe:Compared with the model mice,the expression of IL-6 and IL-10 gene of the inflammatory factor NF-??B,TNF-?,IL-6 and IL-10 in the three intervention groups(the difference of RNA-seq technique in the previous study group)had different changes in the liver tissue(P<0.05,P<0.01),The mRNA expression of inflammatory cytokines in the simple intervention group was significantly different from that in the combined intervention group(P<0.05,P<0.01).The mRNA expression of TNF-?,IL-6 was the highest in the model group,which was significantly different from that in the aerobic exercise group and the flavone group(P<0.05),while the lowest expression of TNF-?IL-6 mRNA was found in the exercise+flavone group(P<0.01).(5)The positive trend of TNF-? gene expression in liver and blood tissues was found by correlation analysis,and the expression of TNF-? in both liver and blood tissues was statistically significant(P<0.01).Conclusion:1.long-term aerobic exercise and black fruit wolfberry flavone have an intervention effect on alcoholic liver injury in mice,which can regulate the changes of inflammatory cytokines in liver injury mice,and its expression level is related to the occurrence and development of inflammatory process of alcoholic fatty liver.Its possible mechanism is to reduce the inflammatory cytokines related to TNF-? and IL-6 in NF-?B signaling pathway,increase the content of IL-10 anti inflammatory cytokines and alleviate the inflammatory symptoms of liver,to effect of km mice on the inflammation of liver induced by alcohol.2.The test of cyclic RNA is an ideal method for obtaining molecular markers by non invasive method.As an important sign of alcoholic liver injury screening diagnosis and observation and prognosis monitoring,it has a good prospect of clinical application.