基于响应面法优化牛蒡子多糖提取工艺及对细胞抗炎作用研究

Optimization of extraction process of Arctii Fructus polysaccharide based on response surface method and its anti-inflammatory effect on cells

为了确定提取牛蒡子多糖的最佳工艺条件,并探究其抗炎作用。通过响应面法考察不同料液比,提取时间和提取温度对牛蒡子多糖提取率的影响;使用ELISA试剂盒测定不同RAW 264.7细胞处理组肿瘤坏死因子α(TNF-α)和白介素-6(IL-6)的水平;经Western blot法检测髓样分化因子88(MyD88)、Toll样受体4(TLR4)和核转录因子-κB(NF-κB)的蛋白表达水平。结果显示牛蒡子多糖最优提取工艺条件为料液比1∶15(g/mL)、提取时间3 h、提取温度80℃,此时提取率为7.19%;牛蒡子多糖组对TNF-α和IL-6的分泌均有抑制作用并通过调控相关通路上的蛋白表达,起到免疫调控作用。该研究可为牛蒡子新药用成分的开发和应用奠定基础。

The aim of this paper was to determine the optimum extraction conditions of Arctii Fructus polysaccharide and to explore the anti-inflammatory effect of its solution.The effects of different solid-liquid ratio,extraction time and extraction temperature on the extraction yield of Arctii Fructus polysaccharide were investigated by response surface method.The levels of tumor necrosis factorα(TNF-α)and interleukin-6(IL-6)in different RAW 264.7 cell treatment groups were determined by ELISA kit.The protein expression levels of myeloid differentiation factor 88(MyD88),Toll-like receptor 4(TLR4)and nuclear transcription factor-κB(NF-κB)were detected by Western blot.The results showed that the optimal extraction conditions were solid-liquid ratio 1∶15(g/mL),extraction time 3 h,extraction temperature 80℃,and the extraction yield was 7.19%.Arctii Fructus polysaccharide groups inhibited the secretion of TNF-αand IL-6,and played an immunoregulatory role by regulating the expression of proteins in related pathways.This study can lay a foundation for the development and application of new drug ingredients of Arctii Fructus.