裸藻β-1,3-葡聚糖提高斑马鱼及细胞免疫活性研究  

Euglena gracilisβ-1,3-Glucan Enhancing Zebrafish and Cellular Immune Activities

作　　者：康佳 张樱馨 邵亚会 杨文龙 耿耘[2,3] 宣宁 吕和鑫[1,4] 陈高 Kang Jia;Zhang Yingxin;Shao Yahui;Yang Wenlong;Geng Yun;Xuan Ning;Lyu Hexin;Chen Gao(College of Bioengineering,Tianjin University of Science and Technology,Tianjin 300457,China;Institute of Crop Germplasm Resources,Shandong Academy of Agricultural Sciences,Jinan 250100,China;National Key Laboratory of Efficient Utilization of Nutrient Resources,Jinan 250100,China;Shandong Nameida Biological Technology Co.,Ltd.,Jinan 250132,China)

机构地区：[1]天津科技大学生物工程学院,天津300457 [2]山东省农业科学院农作物种质资源研究所,山东济南250100 [3]养分资源高效利用全国重点实验室,山东济南250100 [4]山东纳美达生物科技有限公司,山东济南250132

出　　处：《山东农业科学》2025年第2期158-164,共7页Shandong Agricultural Sciences

基　　金：国家自然科学基金项目(32170396,32100016);山东省自然科学基金项目(ZR2022QC154,ZR2023QC149);山东省科技型中小企业创新能力提升工程项目(2023TSGC0204)。

摘　　要：本研究采用斑马鱼模型和细胞模型,比较裸藻粉样品A(纤细裸藻β-1,3-葡聚糖)、样品B(纤细裸藻粉)和样品C(商业纤细裸藻粉)的生物活性差异。首先,使用野生型斑马鱼AB品系,施加不同剂量的待测样品,并观察其对斑马鱼幼鱼存活率的影响。确定样品A对斑马鱼幼鱼的安全剂量为≤400μg/mL,样品B和样品C的安全剂量均为≤200μg/mL。然后,使用免疫细胞转基因斑马鱼品系,在受精卵发育至24 hpf(受精后24 h)时,以100μg/mL的长春瑞滨溶液构建免疫抑制模型,24 h后观察并统计斑马鱼尾部免疫细胞数目。结果显示,50μg/mL和100μg/mL的样品A,以及100μg/mL的样品B和样品C显著增加斑马鱼免疫细胞数量。接着,以巨噬细胞系Raw 264.7为对象,检测样品对免疫细胞增殖的影响,确定各样品的安全剂量均为≤200μg/mL。最后,在安全剂量下,测定药物在脂多糖(LPS)处理后细胞培养液中一氧化氮(NO)含量,以确定各样品在免疫调节(抗炎作用)中的效果。结果样品A在50μg/mL和100μg/mL剂量下,以及样品B和样品C在100μg/mL剂量下均明显降低细胞培养液中的NO含量,表明各样品均具有免疫调节作用,且样品A的效果优于样品B和样品C。本研究结果有助于筛选出具有较高免疫增强活性的纤细裸藻粉样品,可为开发新型免疫增强剂提供科学指导。This study utilized zebrafish model and cellular model to evaluate the bioactivity differences among three different Euglena gracilis powder samples:Sample A(E.gracilisβ-1,3-glucan),Sample B(E.gracilis powder),and Sample C(commercial E.gracilis powder).First,the wild-type AB zebrafish were exposed to various doses of the test samples to assess their effects on larval survival rates.The safe doses were determined to be≤400μg/mL for Sample A,and≤200μg/mL for both Sample B and Sample C.Next,the transgenic zebrafish with labeled immune cells were used to establish an immunosuppressive model.This was done by treating fertilized eggs at 24 h after post-fertilization(hpf)with 100μg/mL vinorelbine,and the number of immune cells in the tail regions of the zebrafish were counted 24 h later.The results showed that Sample A at 50μg/mL and 100μg/mL,as well as Sample B and Sample C at 100μg/mL,significantly increased the number of immune cells in zebrafish.Subsequently,the macrophage cell line Raw 264.7 was employed to investigate the effects of the samples on immune cell proliferation,revealing that the safe dose for all samples was≤200μg/mL.Finally,at the safe doses,the nitric oxide(NO)content in the culture medium was measured after lipopolysaccharide(LPS)treatment to assess the anti-inflammatory immunomodulatory effects of the samples.The results indicated that Sample A at 50μg/mL and 100μg/mL and Samples B and C at 100μg/mL significantly reduced NO level in the Raw 264.7 cell culture medium,suggesting that all samples exhibited immunomodulatory effects.Notably,Sample A showed superior efficacy compared to both Sample B and Sample C.These findings contributed to the identification of E.gracilis powder samples with potent immune-enhancing activity,offering valuable insights for the development of novel immunoenhancers.

关 键 词：纤细裸藻 Β-1 3-葡聚糖 斑马鱼 免疫活性 

分 类 号：TS201.4[轻工技术与工程—食品科学] Q952[轻工技术与工程—食品科学与工程]