首页|蓝莓渣多酚对肝星状细胞的增殖与自噬作用

蓝莓渣多酚对肝星状细胞的增殖与自噬作用

Effects of Blueberry Pomace Polyphenols on Proliferation and Autophagy of Hepatic Stellate Cells

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为了提高蓝莓全资源利用率,降低其加工副产物对环境的危害.通过高效液相色谱串联质谱法(HPLC-MS/MS)分析超高压提取、超声提取和溶剂提取技术对蓝莓渣多酚(BRP)组分及含量的影响.以活化的HSC-T6细胞为模型,通过噻唑蓝法和乳酸脱氢酶法检测BRP对HSC-T6细胞存活率和细胞毒性的影响,透射电镜和吖啶橙染色法观察BRP对HSC-T6细胞自噬的诱导作用,蛋白免疫印迹法(Western blot)检测BRP对HSC-T6细胞自噬相关蛋白的表达量,酶联免疫法(ELISA)分析HSC-T6细胞的纤维化程度.结果表明:BRP中有14种组分,3种提取方法中的矢车菊素-3-O-葡萄糖苷占比最高,分别为57.530%,58.703%,54.830%.3种提取方法所得BRP的组分种类相同,而含量存在较大差异.超高压提取法所得BRP含量显著高于常规溶剂法和超声辅助提取法(P<0.05).BRP能显著抑制HSC-T6细胞增殖(P<0.05),且在一定浓度范围呈现时间-剂量效应关系;通过上调自噬相关蛋白Beclin-1和LC3-Ⅱ表达,下调p62蛋白表达,诱导细胞发生自噬;同时BRP显著下调HSC-T6细胞中COL-Ⅰ、COL-Ⅲ、TNF-β1的分泌量(P<0.05).结论:超高压萃取的BRP组分总含量最高,生物活性功能最强,BRP可通过抑制活化的HSC-T6细胞增殖,诱导细胞自噬从而发挥其抗纤维化效果.
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.

blueberry residue phenolicsultra-high pressureHSC-T6 cellsproliferationautophagy

马剑、赖光荣、胡一阳、贾飞鸿、马艳弘、刘海燕

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江苏省农业科学院农产品加工研究所 南京 210014

克孜勒苏柯尔克孜自治州林业工作管理站 新疆阿图什 845350

南京市溧水区供销合作总社 南京 211200

蓝莓渣多酚 超高压 HSC-T6细胞 增殖 自噬

新疆自治区重点研发计划项目

2023B02028

2024

中国食品学报
中国食品科学技术学会

中国食品学报

CSTPCD北大核心EI
影响因子:1.079
ISSN:1009-7848
年,卷(期):2024.24(9)
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