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模拟失重对白假丝酵母菌增殖和毒性的影响
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“重大新药创制”科技重大专项(2015ZX09J15102);1226工程卫生专业重大项目(AWS16J018)


Effects of simulated microgravity on proliferation and toxicity of Candida albicans
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    摘要:

    【背景】近年来研究发现,失重条件可对一些致病微生物的增殖和毒性产生影响,白假丝酵母菌(Candida albicans)是典型的条件性致病真菌,在太空环境和人体中普遍存在,研究失重条件下白假丝酵母菌的增殖和毒性意义重大。【目的】利用旋转细胞培养系统(Rotary cell culture system,RCCS)模拟失重环境对白假丝酵母菌进行连续传代培养,检测模拟失重环境对白假丝酵母菌增殖情况、毒性以及基因表达的变化。【方法】将白假丝酵母菌接种在旋转生物反应器(High aspect rotating vessel,HARV)中,利用旋转细胞培养系统连续传代培养14 d,然后对菌株进行增殖速率测定、不同pH条件下增殖能力测定、生物膜相对形成能力测定和细胞毒性和动物毒力测定;利用转录组测序技术找出差异表达基因,结合性状分析模拟失重可能对白假丝酵母菌增殖和毒力的影响。【结果】与对照组相比,模拟失重组白假丝酵母菌对数期提前,增殖速率加快,在适宜pH条件下的增殖能力普遍提高,但其生物膜形成能力相对减弱,对LoVo细胞和小鼠的毒性减弱;转录组测序发现,模拟失重组共有280个基因表达差异达1.5倍以上(P<0.05),其中248个上调、32个下调。差异基因经基因功能注释(Gene ontology,GO)和京都基因及基因组百科全书(Kyoto encyclopedia of genes and genomes,KEGG)富集分析发现,相关胞膜形成及细胞分裂基因表达上调,生物膜形成、细胞黏附及共生粘连宿主基因表达下调。【结论】模拟失重环境可引起白假丝酵母菌增殖和毒性水平发生变化,相关改变可为研究失重环境对微生物的影响提供参考。

    Abstract:

    [Background] Recent studies have found that microgravity conditions can affect the proliferation and toxicity of some pathogenic microorganisms. Candida albicans is a typical conditional pathogenic fungus, which is ubiquitous in space environment and human body. It is of great significance to study the proliferation and toxicity of C. albicans under microgravity conditions. [Objective] Rotating cell culture system (RCCS) was used to simulate the microgravity environment for continuous subculture of C. albicans. The changes of proliferation, toxicity and gene expression of C. albicans in simulated microgravity environment were detected. [Methods] C. albicans were inoculated in high aspect rotating vessel (HARV) and cultured in RCCS for 14 days. After the culture completed, the proliferation rates of C. albicans were measured. The proliferation abilities were detected under different pH conditions, and the cytotoxicity and animal toxicity were determined. The differentially expressed genes were identified by RNA-seq. [Results] In the simulated microgravity group, the logarithmic phase of C. albicans was earlier and the proliferation rate was faster, the proliferation ability was generally improved under suitable pH conditions, but its relative biolfilm formation and toxicity to LoVo cells and mice was weakened. RNA-seq revealed that 280 genes expressed differentially more than 1.5 times (P<0.05), of which 248 were up-regulated and 32 were down-regulated in simulated microgravity environment. The differential genes were enriched by GO and KEGG, and the gene expressions of cell membrane formation and cell division were up-regulated in simulated microgravity environment , while the gene expressions of biofilm, cell adhesion and symbiotic adhesion host were down-regulated. [Conclusion] Simulated microgravity environment can change the proliferation and toxicity of C. albicans, and the related changes can provide reference for studying on the influence of microgravity environment on microorganisms.

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赵光显,王佳平,刘宇,刘军莲,李正超,王言吉,韩延平,李勇枝. 模拟失重对白假丝酵母菌增殖和毒性的影响[J]. 微生物学通报, 2019, 46(11): 3048-3057

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  • 在线发布日期: 2019-11-05
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