基于转录组学的金黄色葡萄球菌噬菌体安全性评估

doi:10.13560/j.cnki.biotech.bull.1985.2019-0696

摘要/Abstract

摘要： 多重耐药菌的出现迫使我们将噬菌体治疗视为可能的替代治疗方法之一。旨在采用转录测序技术探讨金黄色葡萄球菌噬菌体在小鼠乳腺灌注治疗中的安全及长期使用的条件。选用24只泌乳10 d的昆明雌鼠,随机分成3组,包括健康对照组（T01）,噬菌体VBSM-A1实验组（T02）和噬菌体cocktail实验组（T03）。用非损伤方法经乳头管分别注入不同组分的金黄色葡萄球菌噬菌体悬浊液于小鼠的第4对（腹部）乳腺内。观察小鼠、组织病理学变化以及乳腺组织中TNF-α含量变化;同时,采用转录组测序技术对小鼠乳腺组织进行分析。结果显示,噬菌体灌注会导致小鼠乳腺呈现轻微炎症反应,但不足以影响小鼠正常生命体征;同时,24 h后噬菌体的数量会明显下降（起始为5×10⁷ PFU/gland;24 h后则分别为10^3.6和10^4.4PFU/gland）,提示在噬菌体给药的方式以及剂量还需要进一步的探索,同时提出了噬菌体和抗生素联合应用以降低噬菌体免疫应答和抗生素耐药性的前景,旨为后续噬菌体的使用从转录组层面提供理论依据和奠定实验基础。

关键词: 转录组测序, 噬菌体疗法, 噬菌体安全性, 小鼠

Abstract: The emergence of multidrug-resistant bacteria has forced us to consider bacteriophage therapy as one of the possible alternative therapies. The aim of this study is to investigate the safety and long-term use condition of Staphylococcus aureus bacteriophage in mammary perfusion therapy in mice by transcriptional sequencing. Twenty-four Kunming female mice lactating for 10 d were randomly divided into three groups,including healthy control group（T01）,bacteriophage VBSM-A1 perfusion group（T02）and bacteriophage cocktail perfusion group（T03）. S. aureus phage suspension of different components was injected into the fourth pair of mammary glands（abdomen）of mice by non-invasive method,and then the mice and the changes of histopathology and TNF-α in the breast tissue were observed. Meanwhile,transcriptome sequencing was used to analyze the mammary gland of mice. The results showed that phage infusion caused slight inflammation in the mammary glands of mice,but did not cause physiological state changes of the mice. At the same time,the number of phages decreased significantly（started：5×10⁷ PFU/gland;24 h latter：10^3.6 and 10^4.4 PFU/gland respectively）during 24 h,suggesting that the way and dosage of phage administration needed further exploration. The prospect of combined application of phages and antibiotics to reduce the phage immune response and antibiotic resistance is proposed,which provides a theoretical and experimental basis for the use of subsequent phages at the transcriptome level.

Key words: transcriptome sequencing, bacteriophage therapy, phage safety, mouse

耿慧君, 邹伟, 崔惠敬, 李晓宇, 王丽丽, 徐永平. 基于转录组学的金黄色葡萄球菌噬菌体安全性评估[J]. 生物技术通报, 2019, 35(12): 64-75.

GENG Hui-jun, ZOU Wei, CUI Hui-jing, LI Xiao-yu, WANG Li-li, XU Yong-ping. Safety Evaluation of Staphylococcus aureus Bacteriophage Based on Transcriptome[J]. Biotechnology Bulletin, 2019, 35(12): 64-75.

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