基于靶向代谢组学研究环形泰勒虫对宿主细胞能量代谢的影响

doi:10.11843/j.issn.0366-6964.2021.012.020

摘要/Abstract

摘要： 环形泰勒虫感染宿主淋巴细胞可使其获得类似肿瘤细胞样的无限增殖能力，而细胞转化现象会伴随虫体的消失而终止，该寄生模式为人们更好地理解环形泰勒虫和细胞相互作用的分子机制提供了一个理想模型。本研究通过靶向代谢组学方法研究环形泰勒虫感染对宿主细胞能量代谢相关代谢物的变化情况，为进一步揭示其转化机制奠定基础。以环形泰勒虫转化细胞为试验材料，设置布帕伐醌（buparvaquone，BW720c）药物处理组、DMSO对照组及空白对照组，观察不同处理对环形泰勒虫感染细胞生长状态的影响；并在72 h时收集细胞样品，利用液相色谱串联质谱（LC-MS/MS）的靶向代谢组学方法对不同处理组的细胞样品进行能量代谢相关代谢物检测，再运用能量数据库对检测到的代谢物变量进行化合物种类鉴别，使用最小判别二乘分析法（partial least squares-discriminant analysis，PLS-DA）筛选差异代谢物。通过对转化细胞的药物试验发现，相对于对照组，试验组在24 h内细胞增殖无明显变化，从36 h开始试验组的细胞生长速度明显低于对照组，在72 h时两组间的活细胞数量相差最大。PLS-DA分析发现，代谢物变量可将药物处理组及DMSO对照组之间聚类区分，并筛选到21种组间差异的代谢物，药物处理组中有8个代谢物上调，13个代谢物下调。经富集分析发现这些代谢物主要是与能量代谢相关的代谢物，如谷氨酰胺、L-乳酸、苹果酸、丙酮酸、异柠檬酸等。同时用相应的试剂盒检测后发现，谷氨酰胺、L-乳酸及丙酮酸的含量在药物处理组和对照组的变化趋势与靶向代谢组学检测的结果一致。本试验通过靶向代谢组学分析环形泰勒虫感染细胞的差异代谢物和代谢途径，说明环形泰勒虫感染可显著影响感染细胞的能量代谢，为进步阐明环形泰勒虫转化细胞的机制提供思路。

关键词: 环形泰勒虫转化细胞, 靶向代谢组学, 能量代谢

Abstract: The Theileria annulata infected host cells obtained unlimited proliferation as tumor cells, and the phenomenon of parasite-dependent transformation is reversible. The T. annulata transformed cells provide an ideal model for better understanding the molecular mechanism of parasite-host cells interactions. In the present study, the targeted metabolomics method was applied out to investigate the changes of metabolites of the transformed cells related with energy metabolism. Aimed to investigate the difference of cells proliferation, Buparvaquone treatment group (BW720c), DMSO control group and blank control group were set. Cell samples were collected at 72 h, and the energy metabolism-related metabolites of the cell samples were detected by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) targeted metabolomics method. Then, the compound types of the metabolites were identified by the energy database. The differential metabolites were screened by the Partial Least Squares-Discriminant Analysis (PLS-DA).With the results of cells number counting, it was found that there has no significant change between the three groups within 24 h. However, the cell growth rate of the experimental group was significantly lower than the other two control groups at 36 h, and the difference between the two groups was largest at 72 h. PLS-DA analysis showed that metabolites variables could be clustered and distinguished between the drug treatment group and DMSO control, and 21 differential metabolites between two groups were screened out. In the drug treatment group, 8 metabolites were up-regulated and 13 metabolites were down-regulated. After enrichment analysis, it was found that these metabolites were mainly related to energy metabolism, such as glutamine, L-lactic, malic, pyruvic acid, and isocitrate. Meanwhile, it was found that the changes of glutamine, L-actic and pyruvic acid contents in the drug treatment group and the control group were consitent with the results of targeted metabolomics detection by using the corresponding kits. The results indicated that T. annulata infection can significantly affect the energy metabolism of the host cells, which lays a foundation to illustrate the mechanisms of cells transformation by T. annulata.

Key words: Theileria annulata transformed cells, targeted metabolomics, energy metabolism

中图分类号:

S855.9

李霞, 李志, 曹天行, 殷宏, 罗建勋, 关贵全, 刘军龙, 赵洪喜. 基于靶向代谢组学研究环形泰勒虫对宿主细胞能量代谢的影响[J]. 畜牧兽医学报, 2021, 52(12): 3535-3545.

LI Xia, LI Zhi, CAO Tianxing, YIN Hong, LUO Jianxun, GUAN Guiquan, LIU Junlong, ZHAO Hongxi. Effects of Theileria annulata Infection on Energy Metabolism of Host Cells Based on Targeted Metabolomics[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3535-3545.

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