阿魏菇多糖提取工艺优化及肝细胞保护作用研究

Optimization of polysaccharide extraction process and hepatocyte protection effect of P. eryngii var. ferulae

  • 摘要:
    目的 以液体发酵阿魏菇菌丝体为原料,利用响应面分析法对其胞内多糖进行提取工艺优化,并探究多糖对WRL68细胞增殖和抗氧化作用。
    方法 以阿魏菇菌株为研究对象,采用单因素试验结合响应面法优化阿魏菇胞肉多糖提取工艺。采用甲基噻唑基四唑法(Methyl Thiazolyl Tetrazolium, MTT)分析其对WRL68细胞损伤的保护作用。
    结果 阿魏菇菌丝体多糖优化后提取工艺为液料比40∶1 mL/g,提取温度68 ℃,超声时间38 min,提取时间2 h,重复提取4次,优化工艺条件下多糖得率为30.8%,相较于未优化之前提高了150.2%。同时测定多糖对1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picrylhydrazyl, DPPH)、·OH和·O2自由基的清除能力,结果表明阿魏菇多糖在体外具有良好的抗氧化活性。此外,600 μg/mL浓度阿魏菇多糖显著促进WRL68细胞增殖,细胞存活率达147.528%,且对H2O2引起的肝细胞氧化损伤具有显著的保护作用。
    结论 优化后的提取工艺可显著提高阿魏菇菌丝体多糖提取率,可为采用液体深层发酵开发和利用阿魏菇多糖提供提取工艺参数;多糖对肝细胞氧化应激损伤具有显著的保护作用,为开发阿魏菇多糖保肝药物提供借鉴。

     

    Abstract:
    Objective The purpose of this study is to optimize the extraction process of intracellular polysaccharides from liquid fermented P. eryngii var. ferulae mycelium by response surface analysis and explore the effects of polysaccharides on the proliferation and antioxidant effects of WRL68 cells.
    Methods Taking the P.eryngii var. ferulae strain as the research object, a single-factor experiment combined with response surface methodology was used to optimize the extraction process of intracellular polysaccharides from P.eryngii var.ferulae. The solid-liquid ratio, temperature, ultrasonic time, extraction time and extraction times were selected for single factor experiments, and the intracellular polysaccharide extraction process of P. eryngii var. ferulae was optimized by the response surface method. Meanwhile, tetrazolium salt method was used to analyze its protective effect on WRL68 cell injury.
    Results Single factor and response surface optimization experiments were carried out on the mycelial polysaccharides of P. eryngii var. ferulae to optimize the polysaccharide extraction process. After optimization, the extraction process of P. eryngii var. ferulae mycelium polysaccharide was 40:1 mL/g liquid ratio, the extraction temperature was 68 ℃, the ultrasonic time was 38 min, the extraction time was 2 h, the extraction was repeated 4 times. Finally, the yield of polysaccharides under optimized process conditions was 30.8%, which was 150.2% higher compared to that before the optimization. At the same time, the scavenging ability of polysaccharides to DPPH, ·OH- and ·O2- free radicals was measured, and the results showed that P. eryngii var. ferulae polysaccharides had good antioxidant activity in vitro. In addition, 600 μg/mL concentration of P. eryngii var. ferulae polysaccharides significantly promoted the proliferation of WRL68 cells with a cell survival rate reaching 147.528% and had a significant protective effect on hepatocyte oxidative damage caused by H2O2.
    Conclusion The optimized extraction process can significantly improve the extraction rate of mycelial polysaccharides from P. eryngii var. ferulae and the polysaccharides have a significant protective effect on oxidative stress damage in hepatocytes. This study could provide extraction process parameters for the development and utilization of P. eryngii var. ferulae polysaccharides by liquid deep fermentation and offer experimental reference for the development of hepatoprotective drugs of P. eryngii var. ferulae polysaccharides.

     

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