鹿胎肽对巨噬细胞RAW264.7的免疫调节作用

张凯月; 李春楠; 兰梦; 王亚苹; 尹馨雪; 张辉; 高晓晨

doi:10.13386/j.issn1002-0306.2019110029

鹿胎肽对巨噬细胞RAW264.7的免疫调节作用

doi: 10.13386/j.issn1002-0306.2019110029

长春中医药大学吉林省人参科学研究院, 吉林长春 130117

基金项目:

2019年度吉林省中医药科技项目（2019050）。

详细信息

作者简介:
张凯月(1996-),女,硕士研究生,研究方向:中药有效成分与应用开发研究,E-mail:zky960523@163.com。

通讯作者:
张辉(1958-),男,博士,教授,研究方向:中药化学,E-mail:zhanghui-8080@163.com

高晓晨(1986-),女,硕士,主管药师,研究方向:中药有效成分的开发与利用,E-mail:gao_xiaochen@hotmail.com。

中图分类号: TS251.2
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出版历程
- 收稿日期: 2019-11-05

Immunomodulatory Effects of Peptides from Deer Fetus on RAW264.7 Macrophages

Jilin Ginseng Academy of Changchun University of Chinese Medicine, Changchun 130117, China

摘要

摘要: 本实验从鹿胎中提取鹿胎肽（DFP），通过体外实验研究其对巨噬细胞RAW264.7的免疫调节作用。采用噻唑蓝（MTT）实验检测超滤组分各肽段的增殖活性，筛选增殖活性高的组分。选用中性红吞噬实验、酶联免疫吸附检测法测定DFP对RAW264.7细胞吞噬指数、一氧化氮（NO）浓度及细胞因子如白介素-1β（interleukin-1β，IL-1β）、IL-6和肿瘤坏死因子子-α（tumor necrosis factor-α，TNF-α）的分泌能力，同时还研究了DFP对巨噬细胞各周期分布的影响。结果表明，分子质量小于1 kDa的DFP-5对巨噬细胞RAW264.7作用后，可显著（P<0.05）提高巨噬细胞的增殖活性、吞噬活性、NO释放量以及细胞因子的分泌水平，并且在DFP-5质量浓度为25 μg/mL时效果最好。细胞周期结果表明，DFP-5能够促进RAW264.7细胞的生长，且主要作用在G0/G1期。以上结果表明，鹿胎肽具有增强免疫能力的潜在作用。
- 鹿胎肽 /
- 巨噬细胞RAW264.7 /
- 免疫调节作用
Abstract: Deer fetus peptides(DFP)was extracted from deer fetus and its immunomodulatory activities in macrophage RAW264.7 was evaluated in vitro. The proliferative activity of each peptide segment in ultrafiltration fraction was determined by methyl thiazolyltetrazolium(MTT)assay,and the components with high proliferative activity were screened. To further evaluate its immunomodulatory activity,the effects of DFP on the phagocytic activity,nitric oxide(NO)and cytokines secretion of RAW264.7 macrophages were detected by inverted microscope observation,neutral red phagocytosis assay and enzyme-linked immunosorbent assay(ELISA),respectively. The effects on the cell cycle distribution of macrophages were also observed. The results showed that DFP-5,with molecula mass less than 1 kDa,could significantly(P<0.05)increase cytokines in RAW264.7 such as nitric oxide(NO),interleukin-1β(IL-1β),interleukin-6(IL-6)and the tumor necrosis factor-α(TNF-α). When the concentration of DFP was 25 μg/mL,the secretion of these factors was the closest to that of the positive control group(LPS). Cell cycle results showed that DFP could promote the proliferation of RAW264.7 cells and the effect was mainly at the G0/G1 phase. All these results indicated that the deer fetus peptide had the potential to activate macrophages and enhance immune function.
- deer fetus peptide /
- RAW264.7 macrophages /
- immunomodulatory effects

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参考文献(24)

[1]	Babini E,Tagliazucchi D,Martini S,et al. LC-ESI-QTOF-MS identification of novel antioxidant peptides obtained by enzymatic and microbial hydrolysis of vegetable proteins[J].Food Chemistry,2017,228:186-196.
[2]	Barros J S,Baptista M G,Bairos V A.Human chorionic gonadotropin in human placentas from normal and preeclamptic pregnancies[J].Archives of Gynecology and Obstetrics,2002,266(2):67-71.
[3]	夏园,乔建林,曾令宇.组织固有巨噬细胞的研究进展[J].细胞与分子免疫学杂志,2016,32(7):996-999.
[4]	Li Y,Meng T T,Hao N,et al. Immune regulation mechanism of Astragaloside IV on RAW264.7 cells through activating the NF-κB/MAPK signaling pathway[J].International Immunopharmacology,2017,49:38-49.
[5]	邓志程,张迪,吉宏武,等.马氏珠母贝免疫活性肽的纯化与鉴定[J].广东海洋大学学报,2017,37(4):78-86.
[6]	艾金霞,李明成,夏薇,等.鹿胎DNA检测试剂盒的研制与评价[J].中国药学杂志,2018,53(4):306-311.
[7]	刘芳.鹿胎多肽的制备、抗氧化性研究及其微胶囊产品开发[D].长春:吉林大学,2016.
[8]	赵国财,苏禹刚.鹿胎制剂对老年雌性大鼠免疫指标的影响[J].畜牧与饲料科学,2014,35(1):29-30.
[9]	刘朋,邱芳萍.梅花鹿胎衣中蛋白质的提取分离[J].现代食品科技,2006,22(4):153-155.
[10]	韩广金,杨桂芹,邹兴淮.不同鹿胎及胎盘制剂对老年雄性大鼠免疫机能影响的研究[J].畜牧与兽医,2006,38(1):8-10.
[11]	张永芳,原媛.微波萃取-考马斯亮蓝法提取大豆蛋白的工艺研究[J].食品工业,2018,39(9):44-48.
[12]	唐开永,周鸿翔,田娅玲,等.低浓度小分子多肽含量测定方法的比较研究[J].食品研究与开发,2018,39(16):144-148.
[13]	Deng C,Shang J Y,Fu H T,et al. Mechanism of the immunostimulatory activity by a polysaccharide from Dictyophora indusiata[J].International Journal of Biological Macromolecules,2016,91:752-759.
[14]	张妍,林昌岫,邵玉健,等.轮叶党参粗多糖对体外培养小鼠脾淋巴细胞及RAW 264.7细胞的免疫活性[J].食品工业科技,2018,39(12):311-315.
[15]	苏娣,张芸,戴竹青,等.毛蕊花糖对巨噬细胞RAW264.7的免疫调节作用[J].食品科学,2013,34(7):250-253.
[16]	叶盛旺,杨最素,李维,等.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.
[17]	Yu J,Xie J,Mao X J,et al. Hepatoxicity of major constituents and extractions of Radix polygoni multiflori and Radix polygoni multiflori praeparata[J].Journal of Ethnopharmacology,2011,137(3):1291-1299.
[18]	苗月,任桂红,甄东,等.蛹虫草多糖调节小鼠巨噬细胞RAW264.7免疫活性的分子机制[J].食品科学,2019,40(9):188-194.
[19]	Lee J Y,Kim Y J,Kim H J,et al. Immunostimulatory effect of laminarin on RAW 264.7 mouse macrophages[J].Molecules,2012,17(5):5404-5411.
[20]	Gavagnin E,Ford M J,Mort R L,et al. The invasion speed of cell migration models with realistic cell cycle time distributions[J].Journal of Theoretical Biology,2019,481:91-99.
[21]	包鸿慧,侯风蒙,周睿,等.真姬菇多糖HMP的分子特性及抗癌免疫活性[J].食品科技,2013,38(6):205-209.
[22]	杨闯.生物活性肽在营养保健中的应用[J].食品科学,2003,24(12):153-154.
[23]	Paul A T,Gohil V M,Bhutani K K.Modulating TNF-α signaling with natural products[J].Drug Discovery Today,2006,11(15/16):725-732.
[24]	Günther C,Martini E,Wittkopf N,et al. Caspase-8 regulates TNF-α-induced epithelial necroptosis and terminal ileitis[J].Nature,2011,477(7364):335-339.