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双氢青蒿素对肝癌细胞氨基酸代谢的影响及机制研究
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| 篇名： | 双氢青蒿素对肝癌细胞氨基酸代谢的影响及机制研究 |
| TITLE： | Study on the Effect and Its Mechanism of Dihydroartemisinin on the Metabolism of Amino Acids in Hepatoma Cells |
| 摘要： | 目的：考察双氢青蒿素（DHA）对人肝癌细胞Huh7、BEL-7402中氨基酸类代谢产物的影响，为阐明DHA参与肝癌细胞代谢的调控机制提供理论基础。方法：采用CCK-8法，测定不同浓度DHA（12.5、25、50、100µmol/L）作用24、48、72h后对两种细胞活性的影响。将两种细胞分别分为对照组和给药组（DHA，25µmol/L），加入不含药或含药培养基后培养24h，均平行操作3份。对各组细胞样品进行衍生化处理后，采用气相色谱联用质谱（GC-MS）法检测细胞中氨基酸代谢物含量变化，结合SIMCA-P软件分析和化合物谱库的比对，筛选出两种细胞中的差异代谢物；采用Metaboanalyst4.0软件对差异代谢物进行通路富集分析。结果：与对照组比较，给药组Huh7细胞或BEL-7402细胞中谷氨酰胺、谷胱甘肽、苯丙氨酸、富马酸、牛磺酸等含量均呈下降趋势。上述两种细胞中分别筛选获得28、29个差异代谢物，两者的共同差异代谢物有10个，包括谷氨酰胺、谷胱甘肽、牛磺酸、富马酸、苯丙氨酸等；两者的差异代谢物分别富集到8、6条通路上，其共同富集通路为氨基酸-tRNA生物合成、天冬氨酸-丙氨酸-谷氨酸代谢、氮代谢、苯丙氨酸代谢、戊糖磷酸途径。结论：DHA能明显降低Huh7细胞和BEL-7402细胞的活性，并能显著降低细胞中谷氨酰胺、谷氨酸、谷胱甘肽、苯丙氨酸等的含量；其可能通过影响天冬氨酸-丙氨酸-谷氨酸代谢通路等机制来调控两种细胞的生长。 |
| ABSTRACT： | OBJECTIVE：To investigate the effects of dihydroartemisinin （DHA）on the metabolism of amino acid metabolites in human hepatocellular carcinoma cells Huh 7 and BEL- 7402，and to provide theoretic basis for clarifying the mechanism of DHA regulating the metabolism of hepatocellular carcinoma cells. METHODS ：CCK-8 method was taken to detect the effect of different concentrations of DHA （12.5，25，50，100 µmol/L）treating for 24，48，72 h on the two kinds of cells. Two kinds of cells were divided into control group and administration group （DHA，25 µmol/L），and then cultured with drug-free or drug-containing medium for 24 h，operated in parallel for three times. After derivatization of cell samples in each group ，GC-MS method was used to detect the content of amino acid metabolites ，combined with SIMCA-P software analysis and compound library comparison ，the differential metabolites in two kinds of cells were screened out. The pathway enrichment analysis of differential metabolism was conducted with Metaboanalyst 4.0 software. RESULTS ：Compared with control group ，the contents of glutamine ，glutathione， phenylalanine，fumaric acid and taurine were trending down in Huh 7 or BEL- 7402 cells. There were 28 and 29 differential metabolites obtained from the above two kinds of cells ，and 10 of them were common differential metabolites ，including glutamine，glutathione，taurine，fumaric acid ，phenylalanine，etc. The differential metabolites were enriched in 8 and 6 pathways respectively. The common enrichment pathways were amino acid-tRNA biosynthesis ，aspartate-alanine-glutamate metabolism ， nitrogen metabolism ，phenylalanine metabolism and pentose phosphate pathway ，etc. CONCLUSIONS ：DHA can significantly reduce the activities of Huh 7 cells and BEL- 7402 cells，and the contents of glutamine ，glutamic acid ，glutathione and phenylalanine，etc. It may regulate the growth of the two kinds of cells by influencing the mechanism of aspartic acid- alanine-glutamate metabolic pathway ，etc. |
| 期刊： | 2020年第31卷第02期 |
| 作者： | 王惠国，李丹，孙颖超，李雨桐，韩鑫龙，张静莹，唐玲 |
| AUTHORS： | WANG Huiguo ，LI Dan，SUN Yingchao ，LI Yutong ，HAN Xinlong ，ZHANG Jingying ，TANG Ling |
| 关键字： | 双氢青蒿素；气相色谱-质谱联用技术；肝癌；机制；代谢通路；氨基酸 |
| KEYWORDS： | Dihydroartemisinin；GC-MS；Liver carcinoma；Mechanism；Metabolic pathway ；Amino acid |
| 阅读数： | 380 次 |
| 本月下载数： | 8 次 |
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