Effects of Blueberry Pomace Polyphenols on Proliferation and Autophagy of Hepatic Stellate Cells
In order to improve the utilization rate of blueberry resources and reduce the environmental harm of its pro-cessing by-products.The effects of ultra-high pressure extraction,ultrasonic extraction,and solvent extraction on the composition and content of polyphenol(BRP)in blueberry residue were investigated by high performance liquid chro-matography(HPLC-MS/MS).Activated HSC-T6 cells were used as models to detect the effects of BRP on the survival rate and cytotoxicity of HSC-T6 cells by thiazole blue and lactate dehydrogenase methods.The autophagy induction effect of BRP on HSC-T6 cells was observed by transmission electron microscopy and acridine orange staining.The expression level of autophagy related proteins in HSC-T6 cells was detected by Western blot.The degree of fibrosis in HSC-T6 cells was analyzed by enzyme-linked immunoassay(ELISA).The results showed that there were 14 components in BRP,and cyanidin-3-O-glucoside accounts for the highest proportion among the three extraction methods,which are 57.530％,58.703％and 54.830％respectively.The BRP components obtained by the three extraction methods were the same,but the content was different.The BRP content of ultra-high pressure extraction method was significantly higher than that of conventional solvent method and ultrasound-assisted extraction method(P＜0.05).BRP significantly inhibited the prolifer-ation of HSC-T6 cells(P＜0.05),and showed a time-dose effect relationship within a certain concentration range.Au-tophagy was induced by up-regulating the expression of Beclin-1 and LC3-Ⅱ,and down-regulating the expression of p62.And,BRP significantly decreased the secretion levels of COL-Ⅰ,COL-Ⅲ and TNF-β1 in HSC-T6 cells(P＜0.05).In conclusion,the total content of BRP components extracted by ultrahigh pressure extraction is the highest and the bio-logical activity is the strongest,and BRP could induce autophagy by inhibiting the proliferation of activated HSC-T6 cells,thus exerting its anti-fibrosis effect.
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