基于斑马鱼模型研究天麻多糖的免疫调节作用  

Assessment of immunomodulatory effect of Gastrodiae rhizomapolysaccharides in zebrafish embryo

作　　者：曾易 白钦如 胡瑞雪 王婷 励娜[3] 胡光 ZENG Yi;BAI Qinru;HU Ruixue;WANG Ting;LI Na;HU Guang(School of Pharmacy and Bioengineering,Chongqing University of Technology,Chongqing 400054,China;Center of Gerontology and Geriatrics,West China Hospital,Sichuan University/West China School of Nursing,Sichuan University,Chengdu 610041,China;Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China)

机构地区：[1]重庆理工大学药学与生物工程学院,重庆400054 [2]四川大学华西医院老年医学中心/干部医疗科/四川大学华西护理学院,成都610041 [3]重庆市中药研究院,重庆400065

出　　处：《重庆理工大学学报（自然科学）》2024年第8期281-290,共10页Journal of Chongqing University of Technology：Natural Science

基　　金：重庆市自然科学基金面上项目(CSTB2023NSCQ-MSX0685);重庆市中小学创新人才培养工程项目(CY230914);重庆地方生物医药与大健康产业发展研究智库建设项目。

摘　　要：天麻多糖(Gastrodiae rhizoma polysaccharides, GRP)具有一定的免疫调节和抗炎作用,然而目前对其机理的研究相对较少,也缺乏系统性的评价模型。本研究首次使用斑马鱼模型评估了GRP的安全性和免疫调节作用,为未来GRP成分的研究和开发提供了重要的数据支持。首先从GRP给药后斑马鱼的存活率、心率和卵黄囊水肿情况等方面进行了安全性评估。采用吖啶橙、DAF-FM DA和DCFH-DA荧光探针等方法分别检测了凋亡细胞、一氧化氮(NO)和活性氧(ROS)水平;通过实时定量PCR(RT-qPCR)研究GRP对脂多糖(LPS)诱导的免疫相关蛋白的基因表达的影响;使用中性红和苏丹黑B染色评估GRP对斑马鱼巨噬细胞及中性粒细胞数量的影响。结果表明,GRP抑制了LPS诱导的心率加快、卵黄囊水肿和形态畸形等病理反应,表明GRP对LPS诱导的斑马鱼胚胎毒性具有保护作用。此外,GRP表现出免疫调节作用,呈剂量依赖性地抑制了LPS诱导的NO和ROS的产生以及细胞凋亡。qPCR结果表明,GRP的抗炎作用与C3a和TNF-α的下调以及IL-10翻译的上调有关。此外,GRP还减少了中性粒细胞和巨噬细胞在炎症部位的积聚,体现出了其对LPS诱导的斑马鱼的保护作用。Gastrodiae rhizoma polysaccharides(GRP)have garnered attention primarily for their hepatoprotective and anti-cancer properties,with limited exploration in immunomodulation and anti-inflammation.This study,for the first time,evaluates the safety and immune regulatory effects of GRP by using a zebrafish model,elucidating its mechanism of action and furnishing important data for future GRP components research and development.Safety evaluations encompassing survival rate,heart rate,and yolk sac edema of zebrafish are conducted.Additionally,acridine orange,DAF-FM DA,and DCFH-DA fluorescent probes are employed to detect apoptotic cells,nitric oxide(NO)and reactive oxygen species(ROS)levels respectively.The impact of GRP on the expression of immune genes induced by lipopolysaccharide(LPS)is studied through real-time quantitative PCR(RT-qPCR).The effects of GRP on the quantity of macrophages and neutrophils in zebrafish are assessed by using neutral red and Sudan Black B staining.Our resultsshow co-incubation of GRP inhibits the pathological reactions such as increased heart rate,yolk sac edema,and morphological malformations induced by LPS,demonstrating GRP has a protective effect against LPS-induced toxicity in zebrafish embryos.Furthermore,GRP exhibits dese-dependent immunomodulatory effects,suppressing the production of NO and ROS and cell apoptosis induced by LPS.The results of qPCR indicate the anti-inflammatory action of GRP is associated with the downregulation of C3a and TNF-αand the upregulation of IL-10 translation.Moreover,GRP also reduces the accumulation of neutrophils and macrophages at sites of inflammation,highlighting its protective role against LPS-induced effects in zebrafish.

关 键 词：天麻 多糖 免疫调节 斑马鱼 抗炎 

分 类 号：R962[医药卫生—药理学]