青蛤酶解多肽对RAW264.7巨噬细胞的免疫调节作用

doi:10.7506/spkx1002-6630-20171214-163

食品科学 ›› 2019, Vol. 40 ›› Issue (7): 185-191.doi: 10.7506/spkx1002-6630-20171214-163

青蛤酶解多肽对RAW264.7巨噬细胞的免疫调节作用

叶盛旺，杨最素，李维，唐云平，黄芳芳，张小军，余方苗，丁国芳

1.浙江海洋大学食品与医药学院，浙江省海洋生物医用制品工程技术研究中心，浙江舟山 316022；2.浙江省海洋水产研究所，浙江舟山 316021

出版日期:2019-04-15 发布日期:2019-05-05
基金资助:
舟山市科技局公益项目（2015C31012）；浙江省自然科学基金青年科学基金项目（LQ16H300001）；浙江省教育厅一般项目（Y201534400）

Immunomodulatory Effects of Peptides from Enzymatic Hydrolysate of Cyclina sinensis on RAW264.7 Macrophages

YE Shengwang, YANG Zuisu, LI Wei, TANG Yunping, HUANG Fangfang, ZHANG Xiaojun, YU Fangmiao, DING Guofang

1. Zhejiang Provincial Engineering Technology Research Center of Marine Biomedical Products, School of Food and Medicine, Zhejiang Ocean University, Zhoushan 316022, China; 2. Marine Fisheries Research Institute of Zhejiang Province, Zhoushan 316021, China

Online:2019-04-15 Published:2019-05-05

摘要/Abstract

摘要： 研究酶解法制备的青蛤多肽（Cyclina sinensis peptides，CSP）对RAW264.7巨噬细胞的免疫调节作用。以RAW264.7巨噬细胞相对增殖率为指标，选用胃蛋白酶对青蛤肉进行酶解，酶解液经超滤、冷冻干燥后，采用噻唑蓝（methyl thiazolyltetrazolium，MTT）法筛选出增殖活性最高的组分进行免疫调节作用研究。采用MTT细胞增殖实验、倒置显微镜观察、中性红吞噬实验、硝酸根还原酶法及酶联免疫吸附检测法（enzyme-linked immunosorbent assay，ELISA）测定CSP-5对RAW264.7巨噬细胞的生长与增殖、细胞形态、吞噬能力、一氧化氮（nitric oxide，NO）分泌能力及细胞因子白介素（interleukin，IL）-1β、IL-6和肿瘤坏死因子-α（tumor necrosis factor-α，TNF-α）分泌能力的影响；同时还研究了CSP-5对巨噬细胞各周期分布的影响。结果表明，分子质量小于3 kDa的CSP-5对RAW264.7细胞的生长与增殖具有明显的促进作用，且主要作用于G0/G1期，对细胞吞噬能力、NO分泌能力和细胞因子分泌能力的提高具有显著的促进效果；形态学观察结果显示经CSP-5作用后巨噬细胞体积变大且长出较多伪足。由此可知，CSP-5具有激活巨噬细胞增强机体免疫力的潜在作用。

关键词: 酶解, 青蛤多肽, RAW264.7巨噬细胞, 免疫调节作用

Abstract: An enzymatic protein hydrolysate from Cyclina sinensis was prepared by treatment with pepsin. The hydrolysate was fractionated by ultrafiltration into five fractions (CSP-1–CSP-5). CSP-5 was found to be most effective in promoting cell proliferation as determined by methyl thiazolyltetrazolium (MTT) assay. To further evaluate its immunomodulatory activity, the effects of CSP-5 on the growth and proliferation, morphology, phagocytic activity, nitric oxide (NO) and cytokines secretion of RAW264.7 macrophages were detected by MTT assay, inverted microscope observation, neutral red phagocytosis assay, nitric acid reductase method and enzyme-linked immunosorbent assay (ELISA), respectively. The effects on the cell cycle distribution of macrophages were also observed. Our results showed that CSP-5, with molecular mass less than 3 kDa, could significantly promote the growth and proliferation of RAW264.7 macrophages, mainly at the G0/G1 phase. The cell phagocytosis, and NO and cytokine secretion capacity were also enhanced by CSP-5. Furthermore, morphological observation showed that macrophages became larger with more pseudopods after treatment with CSP-5. It is clear that the peptide obtained in this study has the potential to activate macrophages and enhance immune function.

Key words: enzymatic hydrolysis, Cyclina sinensis peptides, RAW264.7 macrophages, immunomodulatory effects

中图分类号:

TS201.4

叶盛旺，杨最素，李维，唐云平，黄芳芳，张小军，余方苗，丁国芳. 青蛤酶解多肽对RAW264.7巨噬细胞的免疫调节作用[J]. 食品科学, 2019, 40(7): 185-191.

YE Shengwang, YANG Zuisu, LI Wei, TANG Yunping, HUANG Fangfang, ZHANG Xiaojun, YU Fangmiao, DING Guofang. Immunomodulatory Effects of Peptides from Enzymatic Hydrolysate of Cyclina sinensis on RAW264.7 Macrophages[J]. FOOD SCIENCE, 2019, 40(7): 185-191.

[1]	董烨，张益奇，姚洪正，何光喜，戴志远. 预处理方式对鳙鱼骨蛋白酶解液游离氨基酸和挥发性成分的影响[J]. 食品科学, 2021, 42(8): 179-185.
[2]	李伟，叶嘉宜，陈运娇，曹庸. 桉叶多酚提取物体内外抗氧化活性评价[J]. 食品科学, 2021, 42(5): 160-168.
[3]	徐永霞，曲诗瑶，李涛，赵洪雷，冯媛，李学鹏，季广仁，励建荣. 不同蛋白酶对蓝蛤酶解液风味特性的影响[J]. 食品科学, 2021, 42(4): 190-196.
[4]	孙乐常，刘伟峰，林怡晨，赵阿云，张凌晶，翁凌，曹敏杰. 碱性蛋白酶限制性酶解对蓝圆鲹分离蛋白功能特性的影响[J]. 食品科学, 2021, 42(2): 23-29.
[5]	吴永祥，王婷婷，张梦婷，张瑶，郭孝成，孙汉巨，陈向阳. 徽州臭鳜鱼微生物多样性、品质特性及其酶解产物抗氧化能力分析[J]. 食品科学, 2021, 42(2): 128-134.
[6]	王佳蓉，丁阳月，姜云庆，董和亮，王秋野，程建军. 酶法修饰对大豆分离蛋白凝胶性质影响的研究进展[J]. 食品科学, 2021, 42(15): 329-336.
[7]	费烨，王雍，龚仕英，田艳，王筑婷，雷霆雯，张礼林，汪希兰，李红梅. 苦荞清蛋白酶解物对高糖诱导的HepG2细胞胰岛素抵抗的保护作用[J]. 食品科学, 2021, 42(1): 222-227.
[8]	黄瑞杰，李媚，廖安平，蓝平，钟磊，覃琴，蓝丽红，王雪娇，韦丽明，甘兰芳. 酸解法与酶解法调控右旋糖酐分子质量的比较[J]. 食品科学, 2020, 41(6): 43-50.
[9]	陈凡凡，滕飞，韩松，吴长玲，周艳，郭增旺，王中江，李杨. 酶解对豆腐干凝胶结构和理化特性的影响[J]. 食品科学, 2020, 41(5): 23-30.
[10]	高娟，杜佳馨，吴限，杨倩，方东路，郑惠华，赵立艳，胡秋辉. 羊肚菌酶解液制备美拉德反应肉味调味基料[J]. 食品科学, 2020, 41(24): 242-250.
[11]	黄钦钦，田亚平. 条斑紫菜蛋白酶解液α-葡萄糖苷酶和DPP-IV抑制活性的表征及肽成分解析[J]. 食品科学, 2020, 41(24): 110-116.
[12]	王锐，张迪雅，李晔，苏秀榕. 金枪鱼暗色肉酶解优势肽鉴定及其体外抗氧化和血管紧张素转换酶抑制活性分析[J]. 食品科学, 2020, 41(23): 91-99.
[13]	苏娜，伊丽，吉日木图. 驼乳蛋白α-淀粉酶抑制活性肽的制备与鉴定[J]. 食品科学, 2020, 41(22): 148-157.
[14]	苏娜，吉日木图，伊丽. 驼乳肽生物功能特性的研究进展[J]. 食品科学, 2020, 41(21): 321-329.
[15]	李学鹏，刘晏玮，谢晓霞，朱文慧，励建荣，张玉玉，李婷婷，于建洋，牟伟丽. 热预处理对蓝蛤酶解及酶解液呈味特性的影响[J]. 食品科学, 2020, 41(2): 133-140.

青蛤酶解多肽对RAW264.7巨噬细胞的免疫调节作用

Immunomodulatory Effects of Peptides from Enzymatic Hydrolysate of Cyclina sinensis on RAW264.7 Macrophages

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价