| Aronia melanocarpa Elliot is rich in nutrients,such as polyphenols,mineral elements and amino acids,and has high medicinal value.Its active ingredients play a certain role in anti-oxidation,anti-aging,inhibiting tumor growth and regulating immune level.Alcoholic liver disease(ALD)is a liver dysfunction disease caused by long-term drinking.Its pathogenesis is generally fatty liver,hepatitis,liver fibrosis and even liver cancer.ALD is caused by genetic,nutritional and environmental factors,and its specific pathogenesis is very complex.At present,the research on ALD in Aronia melanocarpa is mostly limited to the phenomenon level,lacks systematic mechanism research and efficacy evaluation.Therefore,it is particularly important to explore the mechanism of Aronia melanocarpa on ALD.In order to find the significant differentially expressed genes of ALD,the protective effect and mechanism of Aronia melanocarpa anthocyanins(AMA)on ALD mice were investigated.We searched for ALD gene chip GSE28619 from GEO database as network pharmacology data.GEO2 R pretreatment was performed on GSE28619,and the threshold of significant difference was set as | log2 FC | > 2.A total of 419 ALD significantly differentially expressed genes were screened,including 189 up-regulated genes and 230 down-regulated genes.In order to explore the biological function and interaction relationship of differentially expressed genes,GO function enrichment analysis was carried out,protein interaction network was drawn,core network modules were extracted by MCODE algorithm and KEGG pathway enrichment analysis was carried out.The results showed that the biological functions of differential genes were significantly enriched in extracellular matrix and glycosaminoglycan binding.A total of 11 core network modules were extracted from the protein interaction network,among which MCODE3 had a higher set and contained 11 nodes.The KEGG pathway was mainly enriched in the antiviral-related pathways,focal adhesion signaling pathways,phosphatidylinositol3-kinase-serine/threonine kinase signaling pathway(PI3K-Akt signaling pathway)and so on.There are few studies on the ALD of COL1A1 gene in MCODE3 in the current literature,and the reports on the relationship between AMA and COL1A1 are limited.Therefore,in this study,the protein Collagen I expressed by COL1A1 gene and the four active monomer components of AMA(cyanidin-3-galactoside,cyanidin-3-arabinoside,cyanidin-3-glucoside,cyanidin-3-xyloside)were subjected to computer-simulated molecular docking calculations.The results predicted that the four active monomers of AMA had lower binding free energy with Collagen I protein.It can bind to amino acids near the active site by hydrogen bonding.In summary,we speculate that Collagen I protein and PI3K-Akt signaling pathway may play an important role in the occurrence and development of ALD.In order to investigate the protective effect of AMA on alcoholic liver injury,ALD mouse model was induced by 42 %(v/v)ethanol.The results showed that the body weight of mice was about 19.25 g before modeling,and the body weight of mice in the control group increased by 30.52±1.88 g after modeling.The weight of mice in the Et group increased by 17.88±1.07 g,the body weight of mice in the AMA-H group increased by 24.07±0.92 g.The weight of mice in the model group was lighter than that in the control group,and the weight gain was slow.The intervention of AMA alleviated the slow weight gain of mice caused by alcohol intake,reduced the liver coefficient of mice(P<0.01),and the contents of AST and ALT in serum of mice were decreased(P<0.01).The contents of TC,TG and LDL-C in serum of mice were down-regulated(P<0.01),and the content of HDL-C was reversed(P<0.05).AMA administration decreased the activity of ALDH and increased the activity of GSH-PX(P<0.01).The pathological morphology of liver tissue in mice showed that the liver cords of mice in the model group were disordered,the liver cells were turbid and swollen,and the cytoplasm was loose and balloon-like.The situation in the AMA administration group was improved.In order to further explore the mechanism of AMA on alcoholic liver injury in mice,we used q PCR to detect the expression levels of inflammatory factors COX-2and IL-6 in ALD mice.Western Blot was used to detect the expression of PI3K-Akt signaling pathway,Keap1/HO-1 signaling pathway and upstream and downstream proteins under the intervention of AMA.The results showed that compared with the Et group,the expression of COX-2 and IL-6 inflammatory factors in liver tissue was significantly down-regulated after AMA intervention.Western Blot results showed that AMA could affect the expression of proteins in the downstream PI3K-Akt signaling pathway by down-regulating Collagen I and activating the expression ofα7n ACh R protein,and enhance the phosphorylation level of PI3 K and Akt protein.AMA could inhibit the overexpression of Keap1 protein and reverse the low expression of Nrf2 protein and HO-1 protein.AMA can up-regulate the expression of glycogen synthase kinase GSK-3β and anti-apoptotic protein Bcl-2,and alleviate liver injury caused by alcohol in mice.In summary,AMA can effectively improve the quality of life of ALD mice,inhibit the secretion of pro-inflammatory factors,and alleviate the symptoms of alcoholic liver injury through PI3K-Akt signaling pathway and Keap1/HO-1signaling pathway.This study aims to provide new ideas for the development and clinical application of AMA fruit in the future and provide a theoretical basis for the protection and treatment of alcoholic liver injury. |
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