Protective Effects Of Cyanidin-3-O-glucoside On Non-alcoholic Fatty Liver Disease Liver Exposed To Advanced Glycation End Products In Mice

Objective:Non-alcoholic fatty liver disease(NAFLD) has a high prevalence and may progress to an irreversible nonalcoholic steatohepatitis and fibrosis that are difficult to reverse.Studies have shown that the development of fibrosis in NAFLD is highly correlated with advanced glycation end products(AGEs) produced by food or in vivo metabolic disorders.As an extensive phytochemical,anthocyanins display a good protective effect on various toxic-induced liver.In this study,animal models were used to investigate the protective effects of cyanidin-3-O-glucoside(C3G),the main monomer of anthocyanin,on the liver injury of NAFLD exposed to AGEs.Method:In this study,adult male C57BL/6J mice were selected to establish a fibrosis model of NAFLD mice,and exogenous AGEs were produced by baking AIN-93G feed or methionine choline deficient(MCD) feed at high temperature.All mice were randomly divided into five groups according to body weight,and fed AIN-93G feed(control) after baking;MCD feed(MCD);MCD feed after baking(BMCD);MCD feed after baking + 60 mg/kg gavage dose of C3G(BMCD+LC3G);MCD feed after baking + 120 mg/kg gavage dose of C3G(BMCD+HC3G).The first three groups were given a certain volume of normal saline per day.In addition,the food intake and body weight of the mice were recorded.Mice were sacrificed 5 weeks later to collect samples.Liver pathological changes were observed by H&E staining and liver fibrosis was observed by sirius red staining.Expression levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT)in serum,and liver triglyceride(TG) content were detected by special detection kit.Interleukin 1?(IL-1?),interleukin 1?(IL-1?),tumor necrosis factor ?(TNF-?) and monocyte chemoattractant protein 1(MCP-1) in serum were detected by high-throughput protein chip.In addition,proteomics was used to screen differentially interesting proteins and analyze their functions and pathways.Result:The results of H&E staining showed that there were fat vacuoles and fat accumulation in the liver samples in the MCD group,while AGEs could aggravate steatosis.The addition of C3G significantly reduced the number and size of fat vacuoles in the liver,suggesting that C3G can reduce fat accumulation in NAFLD exposed to AGEs.MCD led to a significant increase in serum AST and ALT levels,and AGEs aggravated the release of AST from liver to blood.The intervention of C3G significantly reduced the levels of AST and ALT,and showed significant hepatoprotective effects.There was no significant fibrosis in NAFLD mice exposed to AGEs,suggesting that the activation of hepatic stellate cells(HSCs)was not high.By proteomic analysis,4577 proteins were identified and 332,76 and 20 differentially expressed proteins were screened out in batches.Through protein function analysis and pathway analysis,C3G can effectively regulate energy metabolism-related pathways,improve inflammation,and show a significant association with mitochondrial damage,extracellular exosome,endoplasmic reticulum,etc.Conclusion:AGEs aggravate fat accumulation and inflammation in NAFLD mice,while C3G can reduce TG content in the liver and inhibit the expression of inflammatory factors.The protective effect of C3G on NAFLD mice exposed to AGEs may be through the regulation of energy metabolism-related pathways,improving inflammation and mitochondrial damage,or by acting on extracellular exosome and endoplasmic reticulum.This study can provide experimental basis of reasonable dietary guidance for NAFLD patients to take less reasonable intake of high-temperature cooking foods and more intake of anthocyanin foods,so as to reduce or even avoid chronic liver disease.