Changbai Mountain Wild Blueberries Regulate Ethanol-induced Hepatocellular Injury Via SIRT1 Signaling Pathway

Objective:Blueberry(BB) fruits contain abundant anthocyanins and polyphenols. In our previous research, we found the good protective effect of BB. Mounting evidence suggests that SIRT1 play a key role in some physiological and pathological processes. In addition, SIRT1 is a critical signaling molecules controlling the pathways of hepatic lipid metabolism. Therefore, In this study we investigate protective effect of SIRT1-mediated Changbai mountain blueberries regulate ethanol-induced hepatocellular injury and its underlying mechanisms.Methods:After BB extract was extracted by freeze drying methods, the content of cyanide and resveratrol in BB extracts were determined by high performance liquid chromatography (HPLC). In vitro, Human Chang liver cells were treated with different blueberry extracts (31.25、62.5、125 μg/mL) and ethanol (50 mM) in the presence for 48 hours. Cell viability was analyzed by Methylthiazolyldiphenyl tetrazolium bromide, protein expressions were assessed by Western blot. In addition, after Chang liver cells were treated with different BB extracts (31.25% 62.5% 125 μg/mL) and ethanol (50 mM) in the presence, the pathological change and SIRT1 expression of ethanol-induced Chang liver cells were respectively observed by using Oil red staining method and immunofluorescence method.Results:By determination of the HPLC results showed that contents of anthocyanin and resveratrol were 12.732 mg/100 g and 2.162 mg/100 g respectively. In vitro results showed that BB extracts effectively decreased Chang liver cell viability. BB extracts could obviously decrease the expression of SREBP-1% Collagen Ⅰ、α-SMA、 P-LKB1 and regulated the ratio of MMP-13/TIMP-1. BB extracts could significantly up-regulate the expression of SIRT1, and BB extracts could also significantly increase the expression of P-AMPK、P-ACC and PPARa through the SIRT1 signaling pathways. Oil red staining results that compare with the normal group, bright red lipid drops increased around nucleus in the model group. Bright red lipid drops decreased gradually around nucleus after treating with BB extracts. Immunofluorescence results showed that compared with the normal group, the red fluorescence of SIRT1 decreased in the model group, and the red fluorescence of SIRT1 increased gradually after treating with BB extracts.Conclusion:BB extracts have the potential value of the prevention and treatment.BB extracts may regulate the ethanol-induced hepatocellular injury by mediating SIRT1.In vitro results showed that BB extracts was a potential pharmacology SIRT1 agonist.