Li Gairu, Xiang Yang, Zhao Jin, Chang Junmin
Department of Pharmacy, Xinjiang Medical University, Urumqi, Xinjiang 830011, P.R. China.
Center for Medical Laboratory, Clinical Medical Research Institute, The First Affiliated Hospital of Xinjiang Medical University, Urumqi, Xinjiang 830054, P.R. China.
Exp Ther Med. 2018 Apr;15(4):3556-3562. doi: 10.3892/etm.2018.5818. Epub 2018 Jan 31.
The immune activities of Saccharum Alhagi polysaccharides (SAP) have been previously studied. The present study aimed to investigate the effects of SAP-1 and SAP-2 on the activity of RAW264.7 mouse macrophages. RAW264.7 cells were treated with 150, 300 and 600 mg/l concentrations of SAP-1 (a 50% alcohol precipitation) and SAP-2 (an 80% alcohol precipitation) or with 10 mg/l lipopolysaccharide. Untreated cells were used as a negative control. An MTT assay was used to detect the proliferation of the cells, and Hoechst 33528 staining was conducted in order to visualize the cell nuclei. Additionally, the Griess method was used to measure nitric oxide (NO) levels. A neutral red uptake assay was performed to determine the phagocytic activity of the macrophages, and ELISAs were performed to detect cytokine levels. Furthermore, reverse transcription-quantitative polymerase chain reaction was used to measure the mRNA expression of certain cytokines. The results demonstrated that SAP increased the proliferative activity and activated the immune function of RAW264.7 cells, and was lacking in cytotoxicity. In addition, SAP-1 exhibited a stronger effect in promoting RAW264.7 cell proliferation than did SAP-2. Furthermore, SAP-1 and SAP-2 significantly increased the level of NO, with the effect of SAP-1 being stronger than that of SAP-2. SAP-1 increased the phagocytic activity of RAW264.7 cells and promoted the secretion of the cytokines interleukin (IL)-1β, IL-2 and tumor necrosis factor (TNF)-α by RAW264.7 cells, with an effect that was stronger than that of SAP-2. Finally, different concentrations of SAP-1 or SAP-2 had distinct effects in upregulating the expression of TNF-α, IL-1β, nuclear factor-κB and inducible nitric oxide synthase mRNA. The results of the present study demonstrate that SAP is capable of enhancing the immune activity of mouse macrophages.
此前已对骆驼刺多糖（SAP）的免疫活性进行过研究。本研究旨在探讨SAP - 1和SAP - 2对RAW264.7小鼠巨噬细胞活性的影响。用浓度为150、300和600 mg/l的SAP - 1（50%乙醇沉淀）和SAP - 2（80%乙醇沉淀）或10 mg/l脂多糖处理RAW264.7细胞。未处理的细胞用作阴性对照。采用MTT法检测细胞增殖情况，进行Hoechst 33528染色以观察细胞核。此外，采用Griess法测量一氧化氮（NO）水平。进行中性红摄取试验以确定巨噬细胞的吞噬活性，并进行酶联免疫吸附测定（ELISA）以检测细胞因子水平。此外，采用逆转录 - 定量聚合酶链反应来测量某些细胞因子的mRNA表达。结果表明，SAP可提高RAW264.7细胞的增殖活性并激活其免疫功能，且无细胞毒性。此外，SAP - 1在促进RAW264.7细胞增殖方面比SAP - 2表现出更强的作用。此外，SAP - 1和SAP - 2显著提高了NO水平，其中SAP - 1的作用强于SAP - 2。SAP - 1增强了RAW264.7细胞的吞噬活性，并促进RAW264.7细胞分泌细胞因子白细胞介素（IL）-1β、IL - 2和肿瘤坏死因子（TNF）-α，其作用强于SAP - 2。最后，不同浓度的SAP - 1或SAP - 2在上调TNF -α、IL - 1β、核因子 -κB和诱导型一氧化氮合酶mRNA表达方面具有不同作用。本研究结果表明，SAP能够增强小鼠巨噬细胞的免疫活性。
