目的 建立大鼠红细胞磷脂的代谢组学判别模型,综合运用多种统计学分析方法寻找知母多糖治疗2型糖尿病的红细胞磷脂药理标志物.方法 以高脂饲料喂饲8周龄的SD大鼠8周后,腹腔注射40 mg/kg链脲佐菌素建立2型糖尿病模型.给药组分别灌胃知母多糖及二甲双胍干预治疗8周后,取血分离红细胞制备红细胞磷脂.采用UPLC/Q-TOF分析大鼠在2型糖尿病状态下以及经知母多糖干预后红细胞磷脂的变化.数据导入Simca P V12软件进行多元统计分析处理,通过R软件的BioMark包以及MetaboAnalyst 3.0软件进行代谢物筛选.结果 建立了4组大鼠红细胞磷脂的偏最小二乘法判别分析(PLS-DA)模型,正离子下模型的R2=0.934,Q2=0.534,负离子下模型的R2=0.914,Q2=0.642,由PLS-DA模型的得分图上可见,在正、负离子模式下,前3个主成分均可充分提取4组差异信息.通过R软件的BioMark包筛选并通过ROC分析排除假阳性结果,最终共筛选出知母多糖药理标志物10个,筛选得到的知母多糖药理标志物在模型组与知母多糖干预组间差异具有统计学意义(P<0.01).结论 通过交叉检验及置换检验证明,模型可以充分提取组间差异,并具有较好的预测能力.筛选得到的知母多糖红细胞磷脂标志物可为后续的作用机制研究提供参考.%Objective To establish a metabonomics discriminant analysis model to discriminate the rats' erythrocyte phospholipids data,and find out the biomarkers of rats' erythrocyte phospholipids between T2DM group and anemaran treatment (DTM) group by several statistical approaches.Methods The 8 week-age SD rats were fed with high fat fodder for 8 weeks,and then intraperitoneally injected 40 mg/kg streptozotocin to replicate T2DM model.DTM group and metformin treatment (S) group rats were administrated anemaran and metformin respectively by intragastric gavage for 8 weeks.Blood was collected to extract erythrocyte phospholipids,and erythrocyte phospholipids were analyzed by UPLC/Q-TOF.The data was imported to Simca P V12 software to process the multivariate statistical analysis,and biomarkers selection was processed by BioMark package in R software and MetaboAnalyst 3.0 software.Results The PLS-DA models of erythrocyte phospholipids of 4 groups' rats were successfully established.The R2 value and Q2 value were 0.934 and 0.534 in positive ion mode,0.914 and 0.642 in negative ion mode,respectively.Through the score plot of PLS-DA model,the first three components could adequately extract the differences between four groups under the positive and negative ion mode.The Biomark package in R software was used and the false positive results were removed by ROC analysis.10 biomarkers of rats' erythrocyte phospholipids were screened between T2DM group and DTM group and the biomarkers have significant differences.Conclusions Base on Cross validation and Permutation test,the PLS-DA models were established and could sufficiently extract the differences between groups.The selected biomarkers could provide a reference for the continued study on the mechanism of Anemaran.
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