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RCCS模拟微重力对人角化细胞代谢组学的影响
作者:李鳌 姜楠 吴莹莹 闫洪锋 孙培鸣 孙宏伟 徐冰心 周金莲 崔彦 
单位:100083  北京 安徽医科大学战略支援部队特色医疗中心(原解放军第306医院临床学院普通外科)(李鳌、崔彦)  100083  北京 战略支援部队特色医疗中心普通外科(姜楠、闫洪峰、孙培鸣、孙宏伟、崔彦) 特种医学实验研究中心(吴莹莹、徐冰心) 病理科(周金莲) 
关键词:模拟失重 旋转式细胞培养系统 HaCaT细胞 代谢组学 创伤与修复 
分类号:R85
出版年,卷(期):页码:2019,44(9):745-750
摘要:

 [摘要]  目的  探讨旋转式细胞培养系统(RCCS)模拟微重力对人永生化角质形成细胞系(HaCaT)细胞代谢组学的影响。方法  应用RCCS建立模拟微重力细胞培养系统,体外培养HaCaT细胞,随机分为模拟微重力组(SMG)和正常重力组(NG),细胞培养123 d后收集样本,进行LC/MS代谢组学分析。结合正交偏最小二乘法(O-PLS-DA)和两样本独立t检验寻找两组细胞的差异代谢物,将差异代谢物输入KEGG数据库,进行代谢通路的构建与功能分析。结果  LC/MS代谢组学分析结果显示,与NG组比较,SMGHaCaT细胞在RCCS模拟微重力环境中培养1 d后共有74种差异代谢物,其中16种表达上调,58种表达下调;2 d后共有89种差异代谢物,其中15种表达上调,74种表达下调;3 d后共有100种差异代谢物,其中23种表达上调,77种表达下调;以表达有统计学差异(VIP1P0.05)49种代谢物作为目标代谢标记物,其中鞘氨醇、谷氨酸、二十二碳五烯酸下调,脱水山梨糖醇等物质表达上调。KEGG分析显示,其涉及的代谢通路有氨基酸代谢、脂质代谢、细胞增殖和凋亡、物质转运、分解代谢、信号转导等。结论  RCCS模拟微重力环境可对角化细胞代谢产生明显影响,主要涉及鞘脂类、谷氨酸等代谢产物及相关信号通路。

 [Abstract]  Objective  To investigate the effects of RCCS simulated microgravity on the metabolism of the human keratinocyte cell line HaCaT. Methods  The rotary cell culture system (RCCS) was used to simulate the microgravity environment, and HaCaT cells were cultured in vitro and divided randomly into simulated microgravity group (SMG) and normal gravity group (NG). The two group HaCaT cells were collected respectively after 1 d, 2 d and 3 d culture, and the samples were analyzed by LC/MS metabolomics. The differential metabolites between the SMG and NG cells were identified with partial least squares discriminant analysis ((O)PLS-DA), and the data were input into the KEGG database for the construction and functional analysis of metabolic pathways. Results  Comparing to NG cells, after 1 d culture, there were 74 different metabolites in SMG cells, among which 16 were up-regulated and 58 were down-regulated; after 2 d culture, there were 89 different metabolites, among which 15 were up-regulated and 74 down-regulated; after 3d culture, there were 100 different metabolites, of which 23 were up-regulated and 77 were down-regulated. The differentially expressed 49 metabolites (VIP>1 and P<0. 05) after 3 d were set as target metabolites, within which the sphingosine, glutamate, and docosapentaenoic acid were down-regulated, and dehydrated sorbitol was up-regulated. KEGG analysis indicated that the metabolic pathways involved were amino acid metabolism, lipid metabolism, cell proliferation and apoptosis, substance transport, catabolism, and signal transduction. Conclusion  RCCS simulated microgravity may have significant impacts on keratinocyte metabolism mainly involving metabolites such as sphingolipids and glutamate as well as the related signaling pathways.

基金项目:
全军试验技术研究计划重点项目(SYFD1500128)
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