Shen Chun-Yan, Yang Li, Jiang Jian-Guo, Zheng Chao-Yang, Zhu Wei
College of Food and Bioengineering, South China University of Technology, Guangzhou, 510640, China.
The Second Affiliated Hospital, Guangzhou University of Chinese Medicine, Guangzhou 510120, China.
Food Funct. 2017 Feb 22;8(2):796-807. doi: 10.1039/c6fo01545j.
The crude polysaccharides of Citrus aurantium L. var. amara Engl (CAVAPs) were extracted and their bioactivities including DPPH radical scavenging activity, cytotoxicity to human breast cancer cells, MCF-7, as well as lung cancer cells, HCC827, and their immune-enhancement activity were evaluated. Results showed that CAVAPs exhibited better immunoenhancement activity compared to the DPPH radical scavenging and anticancer activities. Subsequently, the immune enhancement activity of CAVAPs on RAW264.7 cells was further observed and the results displayed that CAVAPs could significantly stimulate the production of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) in RAW264.7 cells, and promote the mRNA expression levels of inducible nitric oxide synthase (iNOS), TNF-α, interleukin-1 beta (IL-1β), and IL-6. Furthermore, western blot analysis demonstrated that the phosphorylated extracellular signal-regulated kinase (ERK), phosphorylated c-Jun N-terminal kinase (JNK), phosphorylated p38 and phosphorylated p65 were all remarkably increased in CAVAP-treated RAW264.7 cells. All these results indicated that CAVAPs might activate macrophages through the mitogen-activated protein kinase (MAPK) and nuclear factor-kappa B (NF-κB) signaling pathway. Additionally, a three-level-three-factor Box-Behnken design (BBD) was performed to optimize the extraction process of CAVAPs for the purpose of application and further research. The maximum extraction yield reached 4.49 ± 0.25%.
提取了酸橙（Citrus aurantium L. var. amara Engl）的粗多糖（CAVAPs），并评估了它们的生物活性，包括对DPPH自由基的清除活性、对人乳腺癌细胞MCF-7以及肺癌细胞HCC827的细胞毒性，以及它们的免疫增强活性。结果表明，与DPPH自由基清除和抗癌活性相比，CAVAPs表现出更好的免疫增强活性。随后，进一步观察了CAVAPs对RAW264.7细胞的免疫增强活性，结果显示CAVAPs可显著刺激RAW264.7细胞中肿瘤坏死因子-α（TNF-α）和白细胞介素-6（IL-6）的产生，并促进诱导型一氧化氮合酶（iNOS）、TNF-α、白细胞介素-1β（IL-1β）和IL-6的mRNA表达水平。此外，蛋白质免疫印迹分析表明，在经CAVAP处理的RAW264.7细胞中，磷酸化的细胞外信号调节激酶（ERK）、磷酸化的c-Jun氨基末端激酶（JNK）、磷酸化的p38和磷酸化的p65均显著增加。所有这些结果表明，CAVAPs可能通过丝裂原活化蛋白激酶（MAPK）和核因子-κB（NF-κB）信号通路激活巨噬细胞。此外，为了应用和进一步研究，采用三水平三因素Box-Behnken设计（BBD）对CAVAPs的提取工艺进行了优化。最大提取率达到4.49±0.25%。
