蓝莓酒泥粗提物的制备及其生物活性

采用超声波辅助酶法提取制备蓝莓酒泥粗提物,研究粗提物的制备工艺并分析粗提物的抗氧化性及其对大鼠肝星状细胞HSC-T6细胞生长的影响。通过单因素试验和正交试验优化制备工艺,检测粗提物对1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基、·OH的清除能力,并通过四甲基偶氮唑蓝增殖实验、Hoechst33258荧光染色、流式细胞术研究其对HSC-T6细胞增殖与凋亡的影响。结果表明:制备蓝莓酒泥粗提物的最佳超声条件为超声功率500 W、料液比1∶20(g/mL)、超声时间50 min、提取温度60℃,在此条件下多糖和花色苷的提取量分别为14.24 mg/g和6.13 mg/g。所制备的蓝莓酒泥粗提物对DPPH自由基和·OH具有一定的清除效果,并可显著抑制HSC-T6细胞增殖、诱导HSC-T6细胞凋亡,且在一定的质量浓度范围内,呈现出剂量-时间效应关系,表明蓝莓酒泥粗提物具有良好的抗氧化活性,并对HSC-T6细胞生长具有明显的抑制作用,具有潜在的抗肝纤维化能力。通过诱导HSC-T6细胞凋亡是蓝莓酒泥粗提物抑制细胞生长的作用途径之一。

Preparation and Biological Activities of Crude Extract from Blueberry Wine Lees

The study focused on the preparation of crude extract from blueberry wine lees by ultrasound-assisted
enzymatic extraction. The antioxidant activity and growth inhibitory effect on HSC-T6 cells of the crude extract on were
also evaluated. The optimization of the extraction conditions was performed using one-factor-at-a-time and orthogonal
array design. Subsequently, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical and hydroxyl radical scavenging capacities
were measured to evaluate antioxidant activity, and the growth inhibitory effect of crude extracts on HSC-T6 cells
was assessed by methylthiazoletetrazolium (MTT) colorimetric assay, Hoechst 33258 fluorescence staining, and flow
cytometry. The optimal extraction conditions were determined as follows: ultrasonic power, 500 W; solid/liquid ratio,
1:20 (g/mL); ultrasonication time, 50 min; and extraction temperature, 60 ℃. Under these conditions, the extraction rates of
polysaccharides and anthocyanins reached 14.24 and 6.13 mg/g, respectively. The crude extract from blueberry wine lees
showed good antioxidant activity and its inhibitory effect on the proliferation of HSC-T6 cells was found to be dose- and
time-dependent. Therefore, the crude extract has high antioxidant activity and significant anti-hepatic fibrosis activity and it
also can inhibit the growth of HSC-T6 cells in vitro, which is partly attributed to its ability to induce cell apoptosis.