引用本文:	崔美玉,耿丽华,张全斌.转录组和蛋白组分析海带褐藻多糖硫酸酯对RAW 264.7细胞的免疫调节作用[J].海洋科学,2022,46(11):94-106.

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转录组和蛋白组分析海带褐藻多糖硫酸酯对RAW 264.7细胞的免疫调节作用
崔美玉^1,2,3, 耿丽华^1,2, 张全斌^1,2
1.中国科学院海洋研究所, 实验海洋生物学重点实验室, 山东青岛 266071;2.青岛海洋科学与技术试点国家实验室, 海洋生物与生物技术功能实验室, 山东青岛 266237;3.中国科学院大学, 北京 100049

摘要:

海带(Saccharina japonica)是我国的主要经济褐藻,从海带中制备的褐藻多糖硫酸酯具有免疫调节等多种生物活性,但鲜有基于组学的研究报道。本文采用RNA高通量测序(RNA-sequencing,RNA-seq)转录组学技术以及平行反应监测(Parallel Reaction Monitoring,PRM)靶向定量蛋白组学方法,探究海带褐藻多糖硫酸酯作用于小鼠巨噬细胞RAW 264.7后对巨噬细胞功能和通路的影响。结果表明,褐藻多糖硫酸酯(100μg/mL)作用于RAW 264.7细胞后,差异表达基因主要富集在免疫反应、先天性免疫应答等生物过程以及TNF、NF-κB、Toll样受体等信号通路,也可促进RAW 264.7细胞膜蛋白A类清道夫受体(SR-A)中SCARA3和Toll样受体中TLR4的表达。采用流式细胞仪对褐藻多糖硫酸酯作用后SR-A和TLR4的表达量进行蛋白水平的验证,SR-A和TLR4的表达量均有升高,证实了组学结果的可靠性。本研究结果为利用褐藻多糖硫酸酯开发具有免疫调节作用的功能食品或药物提供依据。

关键词: 褐藻多糖硫酸酯巨噬细胞免疫调节转录组蛋白组

DOI：10.11759/hykx20220304004

分类号:R285.5

基金项目:山东省重大科技创新工程项目(2019JZZY010818)

Transcriptomic and proteomic analysis of the immunomodulatory effects of fucoidan extracted from Saccharina japonica in RAW 264.7 cells

CUI Mei-yu^1,2,3, GENG Li-hua^1,2, ZHANG Quan-bin^1,2

1.Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;2.Marine Biology and Biotechnology Laboratory, Pilot National Laboratory for Marine Science and Technology(Qingdao), Qingdao 266237, China;3.University of Chinese Academy of Sciences, Beijing 100049, China

Abstract:

Saccharina japonica is a significantly economical brown seaweed found in China, and fucoidan extracted from S. japonica has various biological effects, including immunomodulatory. However, few reports are based on omics. In this study, RNA-Sequencing (RNA-Seq) transcriptome technique and parallel reaction monitoring (PRM) relative quantitative proteomics method were used to investigate the effects of fucoidan on the functions and pathways of RAW 264.7 cells. The results showed that the differentially expressed genes were enriched in biological processes, such as the immune response, innate immune response, TNF, NF-κB, and toll-like receptor signaling pathways. We observed that fucoidan promoted the expression of the scavenger receptor class A member 3 and toll-like receptor 4 in RAW 264.7 cells. The expression levels of SR-A and TLR4 in fucoidan-treated RAW 264.7 were verified by flow cytometry. This showed that the expression levels of SR-A and TLR4 increased, confirming the omics' results reliability. The results of this study provided a theoretical basis for developing functional food products or drugs with the immunomodulatory effects of fucoidan.

Key words: fucoidan macrophage immunomodulatory transcriptomics proteomics