RCCS模拟微重力环境对人胃黏膜上皮GES-1细胞代谢组学的影响

Effects of Simulated Microgravity Environment by RCCS on Metabolomics of GES-1 Cells in Human Gastric Mucosa

目的探讨旋转式细胞培养系统(rotary cell culture system,RCCS)模拟微重力环境对人胃黏膜上皮GES-1细胞代谢组学的影响。方法通过体外培养人胃黏膜上皮GES-1细胞,选取3~10代处于对数生长期细胞进行实验,随机分为模拟微重力组和正常重力组,细胞分组培养1、3、5 d,随后进行LC/MS非靶向代谢组学分析。应用正交偏最小二乘判别分析和两样本独立t检验,寻找2组细胞的差异代谢物;再将差异代谢物输入KEGG数据库,构建代谢通路并进行功能分析。结果2组GES-1细胞经RCCS培养1 d后有151种差异代谢物,其中113种表达上调,38种表达下调;RCCS培养3 d后有134种差异代谢物,其中109种上调,25种下调;RCCS培养5 d后有154种差异代谢物,其中131种表达上调,23种表达下调。通过Venn图筛选出2组1、3、5 d的共同差异代谢物(变量重要性>1且P<0.05)74种,其中57种表达上调,17种表达下调。KEGG数据库分析发现有具体名称及相关功能的差异代谢物44种,主要包括磷脂、氨基酸、辅助因子及羧酸,主要涉及脂类代谢、氨基酸代谢、辅酶和维生素代谢、神经递质代谢、碳水化合物代谢、细胞增殖和凋亡、肿瘤调控、膜转运和信号传导等信号通路。结论RCCS模拟微重力环境对人胃黏膜上皮GES-1细胞的代谢产生明显影响,主要涉及脂类代谢和膜结构损害、细胞增殖凋亡及肿瘤调控。

Objective To investigate the effects of simulated microgravity(SMG)environment by rotary cell culture system(RCCS)on metabonomics of human gastric epithelial GES-1 cells.Methods Human gastric epithelial GES-1 cells were cultured in vitro.Three to ten generations of cells in logarithmic growth phase were selected and randomly divided into SMG group and normal gravity group(NG group).Cells in different groups were cultured for 1,3,and 5 d,and then subjected to non-targeted LC/MS based metabolomics analysis.Orthogonal partial least squares discriminant analysis(OPLS-DA)and two sample independent t-test were used to find the differential metabolites of the two groups of cells,and then the differential metabolites were input into KEGG database to construct metabolic pathways and conduct functional analysis.Results GES-1 cells in both groups had 151 differential metabolites after 1 d of RCCs culture,113 of which were up-regulated and 38 down-regulated;134 of which were up-regulated and 25 down-regulated after 3 d of RCCS culture;154 of which were up-regulated and 23 down-regulated after 5 d of RCCS culture.Seventy-four kinds of common differential metabolites(variable importance>1 and P<0.05)at 1,3 and 5 d in two groups were screened out by Venn diagram,among which 57 were up-regulated and 17 were down-regulated.KEGG database analysis found that there were 44 differential metabolites with specific names and related functions,mainly including phospholipids,amino acids,cofactors and carboxylic acids,mainly involving lipid metabolism,amino acid metabolism,coenzyme and vitamin metabolism,neurotransmitter metabolism,carbohydrate metabolism,cell proliferation and apoptosis,tumor regulation,membrane transport and signal transduction pathways.Conclusion SMG environment by RCCS can significantly affect the metabolism of GES-1 cells,which mainly involves lipid metabolism,membrane structure damage,cell proliferation and apoptosis,and tumor regulation.