动物内源性多肽CGA-N46对念珠菌细胞增殖的影响

生物技术通报 ›› 2014, Vol. 0 ›› Issue (2): 148-152.

动物内源性多肽CGA-N46对念珠菌细胞增殖的影响

阎晓慧, 李瑞芳, 张慧茹, 尹艳杰, 卢研博, 卢亚丽

河南工业大学生物工程学院，郑州 450001

收稿日期:2013-09-09 出版日期:2014-02-27 发布日期:2014-02-27
作者简介:阎晓慧，女，硕士研究生，研究方向：微生物与生化药学; E-mail：879153058@qq.com
基金资助:
国家自然科学基金项目（31071922）, 河南省重点科技攻关计划项目（112102310325）, 河南工业大学科学研究基金研究生教育创新计划项目（12YJCX51）

Effect of Animal Endogenous Polypeptide CGA-N46 on Cell Proliferation of Candidas

Yan Xiaohui, Li Ruifang, Zhang Huiru, Yin Yanjie, Lu Yanbo, Lu Yali

（College of Bioengineering，Henan University of Technology，Zhengzhou 450001）

Received:2013-09-09 Published:2014-02-27 Online:2014-02-27

摘要/Abstract

摘要： CGA-N46是人嗜铬粒蛋白N端具有抗真菌活性的多肽片段。采用MTT法检测CGA-N46对克柔念珠菌生长的抑制作用，以β-1，3-葡聚糖酶活性检测克柔念珠菌细胞壁的完整性，采用PI检测克柔念珠菌细胞膜通透性，用罗丹明123检测线粒体膜电位。试验结果证实，0.5 mg/mL的CGA-N46能够明显抑制克柔念珠菌的细胞增殖能力；对其细胞壁的完整性及细胞膜的通透性没有影响；CGA-N46能明显降低线粒体膜电位。推测CGA-N46可能通过影响线粒体膜电位来抑制克柔念珠菌的生长。

关键词: CGA-N46, 克柔念珠菌, 抑菌作用, 线粒体膜电位

Abstract: CGA-N46 is an N terminal derived peptide of human Chromogranin A (CGA) that has anti-fungal activities. The growth inhibition effect on Candida krusei was measured with MTT assay. The cell wall integrity of Candida krusei was examined by monitoring β-1,3-glucanase activity. The effect of CGA-N46 on the permeability of Candida krusei membrane was examined using Propidium iodide detection. The effect of CGA-N46 on mitochondrial membrane potential was evaluated through Rh-123 assay. The results of showed 0.5 mg/mL CGA-N46 could significantly inhibit the proliferation of the C. krusei cells. CGA-N46 had no obvious effect on the β -1, 3-glucanase activity. There was no significant influence on the permeability of C. krusei cell membrane; CGA-N46 could significantly decrease the mitochondrial membrane potential. A conclusion can therefore be made that CGA-N46 may inhibit the growth of Candidas by affecting the mitochondrial membrane potential.

阎晓慧, 李瑞芳, 张慧茹, 尹艳杰, 卢研博, 卢亚丽. 动物内源性多肽CGA-N46对念珠菌细胞增殖的影响[J]. 生物技术通报, 2014, 0(2): 148-152.

Yan Xiaohui, Li Ruifang, Zhang Huiru, Yin Yanjie, Lu Yanbo, Lu Yali. Effect of Animal Endogenous Polypeptide CGA-N46 on Cell Proliferation of Candidas[J]. Biotechnology Bulletin, 2014, 0(2): 148-152.

参考文献

[1] Schmidtchen A. The multiple faces of host defence peptides and proteins[J]. Journal of Innate Immunity, 2012, 4（4）：325-326.
[2] Pazgier M, Ericksen B, Ling M, et al. Structural and functional analysis of the pro-domain of human cathelicidin, LL-37[J]. Biochemistry, 2013, 52（9）：1547-1558.
[3] Alba A, Lopez-Abarrategui C, Otero-Gonzalez AJ. Host defense peptides：an alternative as antiinfective and immunomodulatory therapeutics[J]. Biopolymers, 2012, 98（4）：251-267.
[4] Cuenca-Estrella M, Gomez-Lopez A, Cuesta I, et al. Frequency of voriconazole resistance in vitro among Spanish clinical isolates of Candida spp. According to breakpoints by the antifungal subcommittee of the european committee on antimicrobial susceptibility testing[J]. Antimicrobial Agents Chemother, 2011, 55（4）：1794-1797.
[5] Bassetti M, Taramasso L, Nicco E, et al. Epidemiology, species distribution, antifungal susceptibility and outcome of nosocomial candidemia in a tertiary care hospital in Italy[J]. PLoS One, 2011, 6（9）：e24198.
[6] Horn DL, Neofytos D, Anaissie EJ, et al. Epidemiology and outcomes of candidemia in 2019 patients：data from the prospective antifungal therapy alliance registry[J].Clin Infect Dis, 2009, 48（12）：1695-1703.
[7] 李瑞芳, 熊前程.嗜铬颗粒蛋白A研究进展[J].生命科学研
究, 2010, 14（4）：350-354.
[8] 李瑞芳, 张添元, 罗进贤, 等. 嗜铬粒蛋白N区抗真菌活性片段研究[J]. 中山大学学报, 2006, 45（2）：4055 - 4059.
[9] 卢研博. 动物内源性多肽CGA-N46抗菌谱的测定及其对细胞膜的作用[D].郑州：河南工业大学, 2013.
[10] Bolintinean D, Hazrati E, Davis HT, et al. Antimicrobial mechanism of pore-forming protegrin peptides：100 pores to kill E.coli[J]. Peptides, 2010, 31（1）：1-8.
[11] Beatriz M, Tim B, Tanja S, et al. Specific interaction of the unmodified bacteriocin Lactococcin 972 with the cell wall precursor lipid II[J]. Applied and Environmental Microbiology, 2008, 74（15）：4665-4670.
[12] Liu Y, Bao QQ, Zhuang XS, et al. Protective effect of catalpol on lactacysin–induced injury in SH-SY5Y cells[J]. Chin J Pharmacol Toxicol, 2012, 26（1）：58-62.
[13] Bastianetto S, Ramassamy C, Poirier J, et al. Dehydroepiandrost erone（DHEA）protects hippocampal cells from oxidative stress-induced damage[J]. Brain Res Mol Brain Res, 1999, 66：35-41.
[14] 房舒, 王春梅, 关巍, 等. 蛇床子素结构修饰物JS-B对辣椒疫霉作用机制研究[J]. 沈阳农业大学学报, 2009, 40（2）：169-173.
[15] 陈卫, 杭锋, 赵建新, 等. 微波杀菌过程中大肠杆菌与金黄色葡萄球菌细胞膜通透性的改变[J]. 微生物学报, 2007, 47（4）：697-701.
[16] Takeshi Z, Fuminori Y, Kenji S. Lactococcal membrane-permeabilizing antimicrobial peptides[J]. Appl Microbiol Biotechnol, 2010, 88（1）：1-9.
[17] Bensassi F, Gallerne C, Dein OS, et al. Mechanism of Alternariol monomethyl ether-induced mitochondrial apoptosis in human coloncarcinoma cells[J]. Toxicology Dec, 2011, 290（2-3）：231-241.
[18] Zhao JX, Guo FL, Bai DC, et al. Effects of Fuzheng Yiliu Granules on apoptotic rate and mitochondrial membrane potential of hepato-cellular carcinoma cell line H22 from mice[J]. Journal of Integrative Uediuce, 2006, 4（3）：271- 274.
[19] Fatma B, Cindy G, Ossama S, et al. Mechanism of Alternariol monomethyl ether-induced mitochondrial apoptosis in human colon carcinoma cells[J]. Toxicology, 2011, 290：230-240.