非靶向脂质组学在子痫前期预测中的应用

doi:10.3969/j.issn.1673-8640.2020.02.002

摘要/Abstract

摘要：

目的采用非靶向脂质组学寻找能早期预测子痫前期的生物标志物。方法选取子痫前期高危孕妇,根据后期是否发生子痫前期分为子痫前期组（33例）和无子痫前期组（对照组,33例）。采用超高效液相色谱（UPLC）-飞行时间质谱（TOF）联用技术对所有对象的血清样本进行脂质组学检测。采用差异倍数、非参数Wilcoxon秩和检验、主成分分析（PCA）和偏最小二乘判别（PLS-DA）筛选生物标志物,并对定性的生物标志物进行代谢通路和富集分析。采用受试者工作特征（ROC）曲线评价筛选出的代谢物对子痫前期的诊断价值。结果共筛选出42种生物标志物,其中甘氨胆酸、二十二碳五烯酸和磷脂酰肌醇（PI）（16:1（9Z）/16:0）水平在子痫前期组中明显升高,胆固醇酯（CE）[22:4（7Z,10Z,13Z,16Z）]、二酰甘油（DG）[18:0/20:1（11Z）/0:0]水平明显降低。含2种以上生物标志物的代谢通路分别为鞘脂类代谢途径、甘油磷脂代谢途径和初级胆汁酸生物合成途径,主要参与α-亚麻酸和亚麻酸代谢、缩醛磷脂合成、长链饱和脂肪酸的线粒体β氧化、鞘脂代谢及胆汁酸生物合成。ROC曲线分析结果显示,CE[22:4（7Z,10Z,13Z,16Z）]、DG[18:0/20:1（11Z）/0:0]、二十二碳五烯酸、PI[16:1（9Z）/16:0]及甘氨胆酸诊断子痫前期的曲线下面积分别为0.723、0.715、0.678、0.669和0.660。结论非靶向脂质组学筛选出的生物标志物能有效区分子痫前期患者和无子痫前期孕妇。甘油磷脂代谢、初级胆汁酸代谢、α-亚麻酸和亚麻酸代谢、胆碱代谢与子痫前期发病有关。

关键词: 非靶向脂质组学, 子痫前期, 质谱

Abstract:

Objective Using non-targeted lipidomics to identify biomarkers for the early prediction of preeclampsia. Methods Pregnant women identified with high risk preeclampsia factors were enrolled and classified into preeclampsia group（33 cases） and non-preeclampsia group（control group,33 cases） according to whether or not they developed preeclampsia in subsequent follow-up period. Serum samples from all the subjects were analyzed for lipidomics by ultra performance liquid chromatography（UPLC）/quadrupole time-of-flight（TOF） mass spectrometry. Univariate statistical analysis with fold change value,Wilcoxon rank sum test of nonparametric test,principal component analysis（PCA） and partial least square discriminant analysis（PLS-DA） were used for screening metabolites. Differential metabolites were analyzed for metabolic pathways and the degree of enrichment. Receiver operating characteristic（ROC）curve was used to assess the value of screened metabolites in the diagnosis of preeclampsia. Results A total of 42 differential metabolites were recognized,and glycocholic acid,docosapentaenoic acid and phosphatidylinositol（PI）[16:1（9Z）/16:0] were increased,while cholesteryl ester（CE）[22:4（7Z,10Z,13Z,16Z）] and diacylglycerol（DG）[18:0/20:1（11Z）/ 0:0] were decreased in preeclampsia group. The metabolic pathways containing more than 2 differential metabolites were sphingolipid metabolism,glycerolipid metabolism and primary bile acid biosynthesis. Differential metabolites were mainly involved in alpha-linolenic acid and linolenic acid metabolism,plasmalogen synthesis,mitochondrial oxidation of long-chain saturated fatty acids,sphingolipid metabolism and bile acid biosynthesis. ROC curve analysis showed that the areas under ROC curves of CE [22:4（7Z,10Z,13Z,16Z）],DG [18:0/20:1（11Z）/0:0],docosapentaenoic acid,PI [16:1（9Z）/16:0] and glycocholic acid were 0.723,0.715,0.678,0.669 and 0.660,respectively. Conclusions Lipidomics can effectively distinguish preeclampsia or not. Glycerophospholipid metabolism,primary bile acid metabolism,alpha-linolenic acid and linolenic acid metabolism and choline metabolism may contribute to preeclampsia pathogenesis.

Key words: Non-targeted lipidomics, Preeclampsia, Mass spectrometry

中图分类号:

R446.1

曹正, 董莹, 赵晟隆, 王晶, 张春红, 陈陆, 刘京瑞, 沈敏, 翟燕红, 刘晓巍. 非靶向脂质组学在子痫前期预测中的应用[J]. 检验医学, 2020, 35(2): 100-107.

CAO Zheng, DONG Ying, ZHAO Shenglong, WANG Jing, ZHANG Chunhong, CHEN Lu, LIU Jingrui, SHEN Min, ZHAI Yanhong, LIU Xiaowei. Utility of non-targeted lipidomics in predicting preeclampsia[J]. Laboratory Medicine, 2020, 35(2): 100-107.

图/表 8

参考文献 21

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序号	离子化模式	保留时间/min	质荷比	代谢物名称	VIP	差异倍数	P值
1	负离子	0.65	464.299	甘氨胆酸	3.208	2.740	0.033
2	负离子	2.05	329.248	二十二碳五烯酸	3.184	2.441	0.005
3	正离子	10.83	723.605	CE[22:4（7Z,10Z,13Z,16Z）]	2.971	0.611	0.001
4	正离子	11.40	633.582	DG[18:0/20:1（11Z）/0:0]	2.913	0.623	0.002
5	负离子	4.50	807.504	PI[16:1（9Z）/16:0]	2.842	1.575	0.014
6	负离子	6.86	1 207.766	神经节苷脂 GM3 [d18:1/20:0]	2.754	0.752	0.027
7	正离子	7.51	631.567	DG[18:2（9Z,12Z）/20:0/0:0）	2.731	0.783	0.020
8	负离子	8.28	826.641	磷脂酰乙醇胺[20:2（11Z,14Z）/22:0]	2.615	0.751	0.000
9	正离子	11.01	397.382	β-谷甾醇	2.554	0.645	0.005
10	正离子	10.57	381.350	24-亚甲基胆甾醇	2.523	0.704	0.003
11	正离子	11.04	699.605	CE[20:2（6Z,9Z）]	2.396	0.665	0.011
12	正离子	8.41	799.595	磷脂酰胆碱[18:1（11Z）/18:3（9Z,12Z,15Z）]	2.386	0.756	0.002
13	正离子	8.55	620.539	DG[18:0/18:2（9Z,12Z）/0:0]	2.368	1.295	0.007
14	负离子	7.77	733.622	鞘磷脂[d18:0/18:0]	2.345	1.380	0.008
15	负离子	1.76	391.285	鹅去氧胆酸	2.276	0.652	0.046
16	正离子	7.81	573.488	DG[16:1（9Z）/18:2（9Z,12Z）/0:0]	2.273	1.243	0.005
17	正离子	8.26	1 264.839	神经节苷脂GM3 [d18:1/24:0]	2.219	0.726	0.014
18	正离子	8.23	618.523	DG[18:1（9Z）/18:2（9Z,12Z）/0:0]	2.206	1.292	0.006
19	正离子	7.77	658.540	DG[18:2（9Z,12Z）/20:4（5Z,8Z,11Z,14Z）/0:0]	2.176	1.290	0.003
20	正离子	5.77	781.622	鞘磷脂[d18:0/22:2（13Z,16Z）（OH）]	2.171	1.303	0.010
21	负离子	7.51	889.574	PI[18:0/20:2（11Z,14Z）]	2.126	0.799	0.008
22	负离子	1.69	283.264	硬脂酸	2.098	1.238	0.022
23	负离子	8.23	653.496	DG[18:1（9Z）/18:2（9Z,12Z）/0:0）]	2.075	1.297	0.003
24	正离子	8.19	565.543	神经酰胺（d18:1/18:0）	2.062	1.283	0.002
25	负离子	8.61	628.543	神经酰胺（d18:1/20:0）	2.055	1.321	0.018
26	正离子	10.53	1 005.792	乳糖神经酰胺（d18:1/25:0）	2.048	0.730	0.016
27	负离子	8.61	818.629	半乳糖神经酰胺（d18:1/22:0）	2.045	0.779	0.002
28	正离子	1.14	568.338	溶血磷脂酰胆碱[22:6（4Z,7Z,10Z,13Z,16Z,19Z）]	2.037	1.371	0.012
29	负离子	7.91	305.248	8,11,14-二十碳三烯酸	2.026	1.259	0.043
30	正离子	2.74	449.281	三烯生育酚	2.010	1.371	0.010
31	正离子	8.58	842.665	磷脂酰胆碱[18:2（9Z,12Z）/22:0]	1.974	0.815	0.002
32	正离子	11.14	935.805	DG[18:1（9Z）/20:1（11Z）/20:4（5Z,8Z,11Z,14Z）]	1.957	0.790	0.003
33	正离子	5.72	594.523	DG[18:1（9Z）/16:0/0:0]	1.914	1.260	0.009
34	正离子	8.11	566.491	DG[16:0/16:1（9Z）/0:0]	1.912	1.217	0.024
35	正离子	8.23	761.653	鞘磷脂（d18:0/20:0）	1.904	1.244	0.007
36	正离子	8.91	622.555	DG[18:0/18:1（9Z）/0:0]	1.850	1.234	0.031
37	正离子	10.73	645.558	CE[16:1（9Z）]	1.825	1.399	0.041
38	正离子	11.20	632.634	神经酰胺（d18:1/24:0）	1.802	0.821	0.003
39	正离子	8.62	773.652	鞘磷脂（d18:0/22:0）	1.766	1.269	0.011
40	负离子	5.72	835.535	PI[16:1（9Z）/18:0]	1.762	1.235	0.022
41	正离子	1.02	543.333	溶血磷脂酰胆碱[20:4（8Z,11Z,14Z,17Z）]	1.739	1.357	0.040
42	正离子	8.25	891.673	磷脂酰胆碱[22:4（7Z,10Z,13Z,16Z）/22:1（13Z）]	1.632	0.820	0.017

序号	离子化模式	保留时间/min	质荷比	代谢物名称	VIP	差异倍数	P值
1	负离子	0.65	464.299	甘氨胆酸	3.208	2.740	0.033
2	负离子	2.05	329.248	二十二碳五烯酸	3.184	2.441	0.005
3	正离子	10.83	723.605	CE[22:4（7Z,10Z,13Z,16Z）]	2.971	0.611	0.001
4	正离子	11.40	633.582	DG[18:0/20:1（11Z）/0:0]	2.913	0.623	0.002
5	负离子	4.50	807.504	PI[16:1（9Z）/16:0]	2.842	1.575	0.014
6	负离子	6.86	1 207.766	神经节苷脂 GM3 [d18:1/20:0]	2.754	0.752	0.027
7	正离子	7.51	631.567	DG[18:2（9Z,12Z）/20:0/0:0）	2.731	0.783	0.020
8	负离子	8.28	826.641	磷脂酰乙醇胺[20:2（11Z,14Z）/22:0]	2.615	0.751	0.000
9	正离子	11.01	397.382	β-谷甾醇	2.554	0.645	0.005
10	正离子	10.57	381.350	24-亚甲基胆甾醇	2.523	0.704	0.003
11	正离子	11.04	699.605	CE[20:2（6Z,9Z）]	2.396	0.665	0.011
12	正离子	8.41	799.595	磷脂酰胆碱[18:1（11Z）/18:3（9Z,12Z,15Z）]	2.386	0.756	0.002
13	正离子	8.55	620.539	DG[18:0/18:2（9Z,12Z）/0:0]	2.368	1.295	0.007
14	负离子	7.77	733.622	鞘磷脂[d18:0/18:0]	2.345	1.380	0.008
15	负离子	1.76	391.285	鹅去氧胆酸	2.276	0.652	0.046
16	正离子	7.81	573.488	DG[16:1（9Z）/18:2（9Z,12Z）/0:0]	2.273	1.243	0.005
17	正离子	8.26	1 264.839	神经节苷脂GM3 [d18:1/24:0]	2.219	0.726	0.014
18	正离子	8.23	618.523	DG[18:1（9Z）/18:2（9Z,12Z）/0:0]	2.206	1.292	0.006
19	正离子	7.77	658.540	DG[18:2（9Z,12Z）/20:4（5Z,8Z,11Z,14Z）/0:0]	2.176	1.290	0.003
20	正离子	5.77	781.622	鞘磷脂[d18:0/22:2（13Z,16Z）（OH）]	2.171	1.303	0.010
21	负离子	7.51	889.574	PI[18:0/20:2（11Z,14Z）]	2.126	0.799	0.008
22	负离子	1.69	283.264	硬脂酸	2.098	1.238	0.022
23	负离子	8.23	653.496	DG[18:1（9Z）/18:2（9Z,12Z）/0:0）]	2.075	1.297	0.003
24	正离子	8.19	565.543	神经酰胺（d18:1/18:0）	2.062	1.283	0.002
25	负离子	8.61	628.543	神经酰胺（d18:1/20:0）	2.055	1.321	0.018
26	正离子	10.53	1 005.792	乳糖神经酰胺（d18:1/25:0）	2.048	0.730	0.016
27	负离子	8.61	818.629	半乳糖神经酰胺（d18:1/22:0）	2.045	0.779	0.002
28	正离子	1.14	568.338	溶血磷脂酰胆碱[22:6（4Z,7Z,10Z,13Z,16Z,19Z）]	2.037	1.371	0.012
29	负离子	7.91	305.248	8,11,14-二十碳三烯酸	2.026	1.259	0.043
30	正离子	2.74	449.281	三烯生育酚	2.010	1.371	0.010
31	正离子	8.58	842.665	磷脂酰胆碱[18:2（9Z,12Z）/22:0]	1.974	0.815	0.002
32	正离子	11.14	935.805	DG[18:1（9Z）/20:1（11Z）/20:4（5Z,8Z,11Z,14Z）]	1.957	0.790	0.003
33	正离子	5.72	594.523	DG[18:1（9Z）/16:0/0:0]	1.914	1.260	0.009
34	正离子	8.11	566.491	DG[16:0/16:1（9Z）/0:0]	1.912	1.217	0.024
35	正离子	8.23	761.653	鞘磷脂（d18:0/20:0）	1.904	1.244	0.007
36	正离子	8.91	622.555	DG[18:0/18:1（9Z）/0:0]	1.850	1.234	0.031
37	正离子	10.73	645.558	CE[16:1（9Z）]	1.825	1.399	0.041
38	正离子	11.20	632.634	神经酰胺（d18:1/24:0）	1.802	0.821	0.003
39	正离子	8.62	773.652	鞘磷脂（d18:0/22:0）	1.766	1.269	0.011
40	负离子	5.72	835.535	PI[16:1（9Z）/18:0]	1.762	1.235	0.022
41	正离子	1.02	543.333	溶血磷脂酰胆碱[20:4（8Z,11Z,14Z,17Z）]	1.739	1.357	0.040
42	正离子	8.25	891.673	磷脂酰胆碱[22:4（7Z,10Z,13Z,16Z）/22:1（13Z）]	1.632	0.820	0.017

代谢通路分析	代谢途径中的生物标志物	-logP值
鞘磷脂代谢途径	神经酰胺、乳糖神经酰胺、半乳糖神经酰胺、鞘磷脂	10.145
甘油磷脂代谢途径	磷脂酰乙醇胺、磷脂酰胆碱、溶血磷脂酰胆碱	5.598
初级胆汁酸合成代谢途径	鹅去氧胆酸、甘氨胆酸	2.869

代谢通路分析	代谢途径中的生物标志物	-logP值
鞘磷脂代谢途径	神经酰胺、乳糖神经酰胺、半乳糖神经酰胺、鞘磷脂	10.145
甘油磷脂代谢途径	磷脂酰乙醇胺、磷脂酰胆碱、溶血磷脂酰胆碱	5.598
初级胆汁酸合成代谢途径	鹅去氧胆酸、甘氨胆酸	2.869

生物标志物	AUC（95%可信区间）	最佳临界值（峰面积）	敏感性/%	特异性/%
甘氨胆酸	0.660（0.533~0.772）	632.98	63.6	69.7
二十二碳五烯酸	0.678（0.551~0.788）	1 159.34	72.7	57.6
CE〔22:4（7Z,10Z,13Z,16Z）〕	0.723（0.599~0.826）	9 372.20	63.6	78.8
DG〔18:0/20:1（11Z）/0:0〕	0.715（0.591~0.820）	34 660.74	75.8	66.7
PI〔16:1（9Z）/16:0〕	0.669（0.542~0.780）	33 824.52	84.8	42.4