Preventive Effect And Mechanism Of Anthocyanins From Aronia Melanocarpa Elliot On Hepatic Fibrosis

Aronia melanocarpa Elliot(Black Chokeberry),its fruit is rich in polyphenols,flavonoids,aromatics and other functional components,in which anthocyanin content is as high as 1%.It was found that anthocyanin in Aronia melanocarpa Elliot(AMA)has strong anti-oxidation,anti-cancer,anti-aging,glucose and lipid-lowering functions in the early experiment of our research group.Hepatic fibrosis(HF)is considered to be one of the most common chronic diseases.The main effector cell is HSC.The expression of TGF-β1 increased with the development of fibrosis,and it is also an effective inducer of stellate cell proliferation and collagen production.In addition,TGF-β1 expression is also related to morphological changes in hepatocytes,such as epithelial mesenchymal transition(EMT).Therefore,it is very important to explore the mechanism of EMT for the development of new and effective treatment of liver diseases.It is of great theoretical and practical significance to destroy the production of TGF-β1 or block the signal transduction with specific small molecules for the generation of effective liver disease treatment methods.In view of the current research reports that polyphenols are closely related to liver disease,we speculate that AMA is of great significance in the field of anti-fibrosis.However,the cellular mechanism of AMA in TGF-β1-induced hepatocyte EMT has not yet been explored.Based on TGF-β / Smad signaling pathway,oxidative stress and inflammatory factors have important effects on liver fibrosis.In this study,TGF-β1-induced HSC-T6 cell model and CCl4-induced liver fibrosis mouse model were used,and foreign aid was applied to interfere with liver fibrosis.Cell and molecular biological techniques were used to evaluate the anti-fibrotic effect of AMA.At the same time,the molecular mechanism of AMA against liver fibrosis was further explored,and its effect on the redox system and inflammation accumulation of liver fibrosis was explored.The mechanism of AMA intervention on liver fibrosis was clarified,which provided scientific theoreticalbasis for the enrichment of liver fibrosis theory and the development and utilization of anthocyanin resources.Methods and results:1)Effect of AMA on TGF-β-induced HSC-T6 cell liver fibrosis and its mechanismHSC-T6 cell model was established by TGF-β1 induction.After 24 hours of in vitro culture of HSC-T6 cells,TGF-β1 and different doses of AMA were added to HSC-T6 cells co-culture of 48 hours.It was detected by liver function indicators,and RT-PCR and Western Blotting were used to explore the mechanism and explore the inhibitory effect of different doses of AMA on HSC-T6 cells.The results show that AMA can effectively inhibit aspartate aminotransferase(AST),alanine aminotransferase(ALT)and other indicators,and reduce the mRNA levels of inflammatory factors such as interleukin-1(IL-1)and interleukin-6(IL-6).And inhibit the expression of α-smooth muscle motor protein(α-SMA),collagen Ⅰ(Colagen Ⅰ)and other proteins.2)Intervention effect of AMA on liver fibrosis in miceCCl4-induced liver fibrosis in mice was accelerated,and different concentrations of AMA(20 mg / Kg and 40 mg / Kg)were administered at the same time.AMA has a certain regulation effect on the body weight of mice,and has a significant inhibitory effect on the abnormal increase of liver coefficient.The results of HE staining and Masson’s trichrome staining showed that AMA had a certain intervention on liver fibrosis in the morphological structure of liver tissue.At the same time,AMA has a significant effect on liver function indexes in the blood and liver tissue of liver fibrosis mice.And improve liver tissue oxidative stress(SOD,GSH-PX)levels.The above results indicate that AMA can alleviate the symptoms of liver fibrosis in a dose-dependent manner.3)Intervention mechanism of AMA on liver fibrosis in miceIn this part,the mechanism of AMA was studied in depth on the basis of the evaluation of its anti-hepatic fibrosis effect in the previous period.Detection of mRNA expression of inflammatory factors(IL-1,IL-6,TNF-α,COX-2)in liver tissue of mice with liver fibrosis by AMA.Studies have shown that AMA can reducethe accumulation of inflammation in liver tissue,and its effect Significantly.At the same time,AMA can also effectively inhibit the expression of TGF-β1,P-Smad2 protein and α-SMA and ColagenⅠ proteins in the TGF-β / Smad signaling pathway during liver fibrosis.The above results indicate that AMA’s inhibition of liver fibrosis may be achieved through the TGF-β / Smad signaling pathway,thereby achieving the effect of delaying liver fibrosis.