| 引用本文:张雪1，2，赵苑伶1，2，陈林珍1，2，刘妍1，2，刘静1，2，赵崇军1，2，刘永刚1，马志强1，2.基于斑马鱼模型探究多花黄精多糖的免疫调节作用[J].世界中医药,2023,(06):. |
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| 基于斑马鱼模型探究多花黄精多糖的免疫调节作用 |
| Immune Function Regulation of Polygonatum cyrtonema Hua Polysaccharides Based on Zebrafish Model |
| 投稿时间：2022-03-29 |
| DOI：10.3969/j.issn.1673-7202.2023.06.004 |
| 中文关键词: 多花黄精 多糖 分离纯化 斑马鱼 免疫调节 中性粒细胞 |
| English Keywords:Polygonatum cyrtonema Hua Polysaccharides Separation and purification Zebrafish Immune regulation Neutrophils |
| 基金项目:国家重点研发计划项目（2018YFC1706500） |
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| 中文摘要: |
| 目的：探讨多花黄精多糖（Polygonatum Cyrtonema Hua Polysaccharides，PCP）及PCP-1对氯霉素所致免疫损伤的保护作用。方法：采用水提醇沉法得到黄精粗多糖，除蛋白脱色素后，得到PCP。利用DEAE-Sepharose Fast Flow层析柱和HW-55F凝胶柱先后对PCP进行洗脱，收集洗脱液，减压浓缩，冷冻干燥后得到纯化的PCP-1。以受精后2 d（2 dpf）的转基因斑马鱼为实验动物，采用不同浓度的氯霉素处理，通过比较斑马鱼尾部中性粒细胞的数目，确定氯霉素的最优造模剂量，建立斑马鱼免疫抑制模型。分别加入不同浓度的PCP和PCP-1孵育24 h后，拍照统计斑马鱼泄殖腔至尾部中性粒细胞数目，观察PCP和PCP-1的免疫调节作用。结果：PCP-1为均一多糖，分子量为3 350 Da。与溶剂对照组比较，氯霉素组的中性粒细胞数目显著减少（P<0.01）；与氯霉素组比较，在实验剂量范围内，PCP和PCP-1均可以增加斑马鱼体内中性粒细胞的数目（P<0.01），且量效关系明显。结论：PCP和PCP-1均具有一定的增强免疫活性及改善免疫损伤的作用。 |
| English Summary: |
| To investigate the protective effects of Polygonatum cyrtonema Hua polysaccharides(PCP) and PCP-1 on immune injury induced by chloramphenicol.Methods:The crude polysaccharides were obtained by water extraction and alcohol precipitation method，and PCP was obtained after protein removal and pigment removal.DEAE-sepharose Fast Flow chromatography column and HW-55F gel column were used to elute PCP.The eluent was collected，condensed under pressure，and freeze-dried to obtain purified PCP-1.Transgenic zebrafish with 2 days of post-fertilization(2 dpf) as experimental animals were treated with different concentrations of chloramphenicol.By comparing the number of neutrophils in the tail of zebrafish，the optimal modeling dose of chloramphenicol was determined，and the immunosuppression model of zebrafish was established.PCP and PCP-1 with different concentrations were added and incubated for 24 h，respectively，and the number of neutrophils from the cloaca to the tail of zebrafish was taken to investigate the immune-enhancing effects of PCP and PCP-1.Results:PCP-1 was a homogenous polysaccharide with a molecular weight of 3 350 Da.As compared with the control group，the number of neutrophils in the chloramphenicol group was significantly reduced(P<0.01).Compared with the chloramphenicol group，both PCP and PCP-1 increased the number of neutrophils in zebrafish within the experimental dose range(P<0.01)，and the dose-response relationship was obvious.Conclusion:Both PCP and PCP-1 can enhance immune activity and ameliorate immune injury to some extent. |
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