开心散多糖提取工艺优化及其体外抗氧化、抗炎活性评价

    Optimization of Extraction Process and Evaluation of in Vitro Antioxidant and Anti-inflammatory Activities of Kai-Xin-San Polysaccharides

    • 摘要:
      目的 优化开心散多糖热水浸提法的提取工艺,并对其体外抗氧化、抗炎活性进行系统评价。
      方法 在单因素试验的基础上,以料液比、提取时间、提取温度为影响因素,多糖得率为评价指标,引入响应面法优化提取工艺;测定多糖对DPPH 自由基、ABTS+自由基、羟基自由基与超氧阴离子自由基的清除率。采用脂多糖诱导并建立体外细胞炎症模型,给予不同浓度开心散多糖进行干预,采用Griess法检测细胞上清液中一氧化氮(NO)的含量,采用ELISA法检测肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素1β(interleukin-1β,IL-1β)和IL-6的含量。
      结果 开心散多糖的最佳提取工艺:料液比1∶20(g·mL−1)、提取时间100 min、提取温度62 ℃,在此条件下多糖得率为(5.734±0.106)%。开心散多糖具有较强的抗氧化活性,其对DPPH 自由基、ABTS+自由基、羟基自由基、超氧阴离子自由基的IC50值分别为0.581、0.212、3.23、1.33 mg·mL−1。细胞实验表明,开心散多糖能抑制巨噬细胞中TNF-α、NO、IL-1β与IL-6的释放(P<0.05或P<0.01),且呈剂量依赖关系。
      结论 本实验建立的开心散多糖提取工艺,方法稳定可行;开心散多糖具有较强的抗氧化活性,且体外抗炎效果显著。

       

      Abstract:
      OBJECTIVE To optimize the extraction process of Kai-Xin-San polysaccharides(KXS-P) by hot water extraction, and to systematically evaluate its in vitro antioxidant and anti-inflammatory activities.
      METHODS On the basis of single factor experiments, response surface methodology was used to optimize the extraction process with the material liquid ratio, extraction time and extraction temperature as influencing factors, and the polysaccharide yield as evaluation indicators. The antioxidant activity of polysaccharides was evaluated according to the clearance rate on DPPH radicals, ABTS+ radicals, hydroxyl radicals, and superoxide anion radicals. The in vitro cellular inflammation model was induced by lipopolysaccharide, and different concentrations of KXS-P were administered for intervention. The content of NO in the cell supernatant was detected using the Griess method, while the levels of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β), and IL-6 were measured using the ELISA method.
      RESULTS The optimal extraction conditions of KXS-P were as follows: material liquid ratio of 1∶20(g·mL−1); extraction time of 100 min; extraction temperature of 62 ℃. The polysaccharide yield was (5.734±0.106)% under the optimal conditions. KXS-P had strong antioxidant activity with IC50 values of 0.581, 0.212, 3.23, 1.33 mg·mL−1 for DPPH, ABTS+, hydroxyl, and superoxide anion radicals, respectively. The results from cellular test showed that KXS-P could inhibit TNF-α, NO, IL-1β, and IL-6 in macrophages(P<0.05 or P<0.01) and the inhibition effect was dose-dependent.
      CONCLUSION The optimized extraction process of KXS-P is stable and feasible. KXS-P have strong antioxidant activity and significant anti-inflammatory effects in vitro.

       

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