液相色谱质谱法对47例非透析慢性肾脏病患者色氨酸-犬尿氨酸通路的分析

doi:10.3877/cma.j.issn.2095-3216.2020.06.002

目的

利用液相色谱-质谱(LC-MS)代谢组学方法探讨慢性肾脏病(chronic kidney disease，CKD)1-4期非透析患者色氨酸(tryptophan, Trp)-犬尿氨酸(kynurenine, Kyn)通路的改变及意义。

方法

2018年3月至2018年6月苏州大学附属第一医院肾内科CKD1～4期非透析患者47例，同期30例健康志愿者作为对照组。应用LC-MS法对受试者血浆进行代谢物分析。应用Human Metabolome Database (HMDB)，MassBank，LipidMaps等数据库匹配不同CKD分期Kyn通路差异代谢物，绘制受试者曲线(receiver operating characteristic, ROC)评估差异代谢物敏感性与特异性。采用Metaboanalyst软件进行代谢通路拓扑分析。以Kyn/Trp比值(KT值)表示Kyn通路限速酶吲哚胺-2，3-双加氧酶(indoleamine-2，3-dioxygenase, IDO)活性。

结果

Kyn在CKD2期即有明显升高(P<0.05)，犬尿喹啉酸(kynurenine acid, KA)在CKD3期明显升高(P<0.01)，二者随CKD分期增加呈上升趋势(P<0.01)。KT值在CKD2期即有升高(P<0.05)，且随CKD分期增加呈上升趋势(P<0.01)。Trp代谢在CKD早期即出现紊乱。

结论

Trp-Kyn通路在CKD早期即增强，Kyn、KA、IDO可能作为CKD早期诊断及进展的生物标志物。

关键词: 慢性肾脏病, 色氨酸-犬尿氨酸通路, 代谢组学, 吲哚胺-2, 3-双加氧酶

Objective

To explore the changes and significance of the tryptophan (Trp)- kynurenine (Kyn) pathway in non-dialysis patients with chronic kidney disease (CKD) stage 1-4 by liquid chromatography-mass spectrometry (LC-MS) metabolomics method.

Methods

From March 2018 to June 2018, 47 non-dialysis patients with CKD stage 1-4 from the Department of Nephrology, First Hospital Affiliated to Suzhou University were recruited, together with 30 healthy volunteers as the control group. LC-MS was used to analyze the blood samples of the subjects. Databases as Human Metabolome Database (HMDB), MassBank, and LipidMaps, etc, were used to match different metabolites of the kynurenine pathway of different CKD stages, and draw the receiver operating characteristic (ROC) curve to evaluate the sensitivity and specificity of the different metabolites. The Metaboanalyst software was used for metabolic pathway topology analysis. The ratio of Kyn to Trp (KT value) was used to express the activity of the indoleamine-2, 3-dioxygenase (IDO), a rate-limiting enzyme in the kynurenine pathway.

Results

In comparison with those of the control, Kyn increased in CKD stage 2 (P<0.05) while kynurenic acid (KA) increased in CKD stage 3 (P<0.01). Both Kyn and KA increased with the progressive loss of renal function (P<0.01). KT value increased in CKD stage 2 (P<0.05), and also increased with the progressive loss of renal function (P<0.01). Disorder of Trp metabolism was observed in the early stage of CKD.

Conclusion

The Trp-Kyn pathway was enhanced in the early stage of CKD. Kyn, KA, and IDO might be used as biomarkers of both CKD early diagnosis and CKD progression.

Key words: Chronic kidney disease, Tryptophan-kynurenine pathway, Metabonomics, Indoleamine-2, 3-dioxygenase

表1 47例非透析CKD患者及30例对照一般临床资料

项目		对照组(n＝30)	CKD1期(n＝13)	CKD2期(n＝11)	CKD3期(n＝11)	CKD4期(n＝12)	P值
基本情况
	年龄(岁)	48±11	47±16	51±21	56±17	58±14	0.232
	性别(男/女)	10/20	5/8	7/4	7/4	7/5	0.223
BMI^a(kg/m²)		1.37±0.07	1.38±0.06	1.36±0.58	1.40±0.04	1.38±0.05	0.755
实验室指标
	肌酐^a(μmol/L)	1.80±0.08	1.74±0.10	1.97±0.10	2.14±0.09	2.37±0.07	<0.001
	白蛋白(g/L)	48(46，49)	36(29，41)	40(37，43)	43(39，44)	41(38，43)	<0.001
	尿素氮^a(mmol/L)	0.67±0.10	0.66±0.14	0.79±0.12	0.95±0.12	1.09±0.16	<0.001
	总胆固醇(mmol/L)	4.5(3.9，5.2)	5.2(4.1，5.6)	4.7(4.5，5.5)	4.5(4.2，5.1)	4.5(3.8，5.2)	0.419
	甘油三酯^a(mmol/L)	0.12±0.23	0.20±0.29	0.16±0.17	0.20±0.11	0.24±0.19	0.493
	血糖(mmol/L)	4.9(4.6，5.2)	4.6(4.5，4.8)	4.6(4.1，4.9)	4.7(4.1，4.8)	4.6(4.3，5.0)	0.086
	血红蛋白(g/L)	140.0±11.4	128.5±17.7	126.6±11.4	121.9±12.8	111.3±17.9	<0.001
	尿酸(μmol/L)	344.5±91.0	379.8±122.7	430.3±59.8	431.2±129.7	420.4±138.4	0.060
临床用药情况
	ACEI类/ARB类	0/30	1/13	2/11	6/11	5/12	-
	前列腺素E1	0/30	0/13	0/11	5/11	6/12	-
	调脂药	0/30	1/13	1/11	6/11	5/12	-
	抗凝药	0/30	1/13	1/11	5/11	6/12	-
	α-酮酸	0/30	1/11	1/11	6/11	6/12	-
	中成类药	0/30	1/13	2/11	6/11	6/12	-

表1 47例非透析CKD患者及30例对照一般临床资料

项目		对照组(n＝30)	CKD1期(n＝13)	CKD2期(n＝11)	CKD3期(n＝11)	CKD4期(n＝12)	P值
基本情况
	年龄(岁)	48±11	47±16	51±21	56±17	58±14	0.232
	性别(男/女)	10/20	5/8	7/4	7/4	7/5	0.223
BMI^a(kg/m²)		1.37±0.07	1.38±0.06	1.36±0.58	1.40±0.04	1.38±0.05	0.755
实验室指标
	肌酐^a(μmol/L)	1.80±0.08	1.74±0.10	1.97±0.10	2.14±0.09	2.37±0.07	<0.001
	白蛋白(g/L)	48(46，49)	36(29，41)	40(37，43)	43(39，44)	41(38，43)	<0.001
	尿素氮^a(mmol/L)	0.67±0.10	0.66±0.14	0.79±0.12	0.95±0.12	1.09±0.16	<0.001
	总胆固醇(mmol/L)	4.5(3.9，5.2)	5.2(4.1，5.6)	4.7(4.5，5.5)	4.5(4.2，5.1)	4.5(3.8，5.2)	0.419
	甘油三酯^a(mmol/L)	0.12±0.23	0.20±0.29	0.16±0.17	0.20±0.11	0.24±0.19	0.493
	血糖(mmol/L)	4.9(4.6，5.2)	4.6(4.5，4.8)	4.6(4.1，4.9)	4.7(4.1，4.8)	4.6(4.3，5.0)	0.086
	血红蛋白(g/L)	140.0±11.4	128.5±17.7	126.6±11.4	121.9±12.8	111.3±17.9	<0.001
	尿酸(μmol/L)	344.5±91.0	379.8±122.7	430.3±59.8	431.2±129.7	420.4±138.4	0.060
临床用药情况
	ACEI类/ARB类	0/30	1/13	2/11	6/11	5/12	-
	前列腺素E1	0/30	0/13	0/11	5/11	6/12	-
	调脂药	0/30	1/13	1/11	6/11	5/12	-
	抗凝药	0/30	1/13	1/11	5/11	6/12	-
	α-酮酸	0/30	1/11	1/11	6/11	6/12	-
	中成类药	0/30	1/13	2/11	6/11	6/12	-

图1 47例非透析CKD患者与30例对照血浆基峰色谱图

图2 47例非透析CKD患者与30例对照血浆样本OPLS-DA图

图3 色氨酸-犬尿氨酸代谢通路

表2 CKD1期患者和对照组色氨酸-犬尿氨酸通路血浆差异代谢物鉴定

	差异代谢物	AUC值	灵敏度	特异度	Youden指数	Ci	Ci2
1	犬尿氨酸	0.524	0.615	0.533	0.148	0.338	0.718
2	犬尿喹啉酸	0.596	0.538	0.700	0.238	0.474	0.756

表2 CKD1期患者和对照组色氨酸-犬尿氨酸通路血浆差异代谢物鉴定

	差异代谢物	AUC值	灵敏度	特异度	Youden指数	Ci	Ci2
1	犬尿氨酸	0.524	0.615	0.533	0.148	0.338	0.718
2	犬尿喹啉酸	0.596	0.538	0.700	0.238	0.474	0.756

图4 色氨酸-犬尿氨酸通路关键代谢物变化趋势

图5 色氨酸代谢通路影响因子

[15]	Mohib K, Wang S, Guan QN, et al. Indoleamine 2，3-dioxygenase expression promotes renal ischemia-reperfusion injury [J]. Am J Physiol Renal Physiol, 2008, 295(1): F226-F234.
[16]	Gargaro M, Vacca C, Massari S, et al. Engagement of nuclear coactivator 7 by 3-hydroxyanthranilic acid enhances activation of aryl hydrocarbon receptor in immunoregulatory dendritic cells [J]. Front Immunol, 2019, 10: 1973.
[17]	Wang CC, Yang CJ, Wu LH, et al. Eicosapentaenoic acid reduces indoleamine 2，3-dioxygenase 1 expression in tumor cells [J]. Int J Med Sci, 2018, 15(12): 1296-1303.
[18]	Zhu X, Li S, Zhang Q, et al. Correlation of increased Th17/Treg cell ratio with endoplasmic reticulum stress in chronic kidney disease [J]. Medicine (Baltimore), 2018, 97(20): e10748.
[19]	Mondanelli G, Iacono A, Carvalho A, et al. Amino acid metabolism as drug target in autoimmune diseases [J]. Autoimmun Rev, 2019, 18(4): 334-348.
[1]	Bikbov B, Purcell C, Levey AS, et al. Global, regional, and national burden of chronic kidney disease, 1990-2017: a systematic analysis for the global burden of disease study 2017 [J]. Lancet, 2020, 395(10225): 709-733.
[2]	Barrios C, Spector TD, Menni C. Blood, urine and faecal metabolite profiles in the study of adult renal disease [J]. Arch Biochem Biophys, 2016, 589: 81-92.
[3]	Gathungu RM, Kautz R, Kristal BS, et al. The integration of LC-MS and NMR for the analysis of low molecular weight trace analytes in complex matrices [J]. Mass Spectrom Rev, 2020, 39(1-2): 35-54.
[4]	Andrassy KM. Comments on 'KDIGO 2012 clinical practice guideline for the evaluation and management of chronic kidney disease’ [J]. Kidney Int, 2013, 84(3): 622-623.
[5]	Sorgdrager F, Naude P, Kema IP, et al. Tryptophan metabolism in inflammaging: from biomarker to therapeutic target [J]. Front Immunol, 2019, 10: 2565.
[6]	Addi T, Dou L, Burtey S. Tryptophan-derived uremic toxins and thrombosis in chronic kidney disease [J]. Toxins (Basel), 2018, 10(10): 412.
[7]	Zinellu A, Sotgia S, Mangoni AA, et al. Impact of cholesterol lowering treatment on plasma kynurenine and tryptophan concentrations in chronic kidney disease: Relationship with oxidative stress improvement [J]. Nutr Metab Cardiovas, 2015, 25(2): 153-159.
[8]	Yoshimura H, Sakai T, Kuwahara Y, et al. Effects of kynurenine metabolites on mesangial cell proliferation and gene expression [J]. Exp Mol Pathol, 2009, 87(1): 70-75.
[9]	Kalaska B, Pawlak K, Domaniewski T, et al. Elevated levels of peripheral kynurenine decrease bone strength in rats with chronic kidney disease [J]. Front Physiol, 2017, 8: 836.
[10]	Bartosiewicz J, Kaminski T, Pawlak K, et al. The activation of the kynurenine pathway in a rat model with renovascular hypertension [J]. Exp Biol Med (Maywood), 2017, 242(7): 750-761.
[11]	Vavrincova-Yaghi D, Seelen MA, Kema IP, et al. Early posttransplant tryptophan metabolism predicts long-term outcome of human kidney transplantation [J]. Transplantation, 2015, 99(8): e97-e104.
[12]	Dschietzig TB, Kellner KH, Sasse K, et al. Plasma kynurenine predicts severity and complications of heart failure and associates with established biochemical and clinical markers of disease [J]. Kidney Blood Press Res, 2019, 44(4): 765-776.
[13]	Boros FA, Vecsei L. Immunomodulatory effects of genetic alterations affecting the kynurenine pathway [J]. Front Immunol, 2019, 10: 2570.
[14]	Schefold JC, Zeden JP, Fotopoulou C, et al. Increased indoleamine 2，3-dioxygenase (IDO) activity and elevated serum levels of tryptophan catabolites in patients with chronic kidney disease: a possible link between chronic inflammation and uraemic symptoms [J]. Nephrol Dial Transplant, 2009, 24(6): 1901-1908.

[1]	何梦媛, 胡鸿保, 谢庆云, 廖冬发, 王维. 股骨头坏死的代谢组学的相关研究进展[J/OL]. 中华关节外科杂志(电子版), 2024, 18(03): 379-382.
[2]	费一鸣, 刘卓, 张丽娟. 组学分析在早产分子机制中的研究现状[J/OL]. 中华妇幼临床医学杂志(电子版), 2024, 20(05): 504-510.
[3]	凌淑洵, 涂玥, 刘思逸. 间充质干细胞在慢性肾脏病研究领域现状和趋势的知识图谱可视化分析[J/OL]. 中华细胞与干细胞杂志(电子版), 2024, 14(02): 73-82.
[4]	中华医学会器官移植学分会, 中国医师协会器官移植医师分会. 中国肝移植受者肾损伤管理临床实践指南（2023版）[J/OL]. 中华肝脏外科手术学电子杂志, 2024, 13(03): 276-288.
[5]	程柏凯, 杨光. 高胰岛素-正葡萄糖钳夹技术评估慢性肾脏病患者胰岛素抵抗的研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(06): 334-339.
[6]	郭俊楠, 林惠, 任艺林, 乔晞. 氨基酸代谢异常在急性肾损伤向慢性肾脏病转变中的作用研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(05): 283-287.
[7]	冯熔熔, 苏晓乐, 王利华. 慢性肾脏病患者并发心血管疾病相关生物标志物研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(05): 273-278.
[8]	王小龙, 吴杰, 段姝伟, 王超卉, 潘娜, 白圆圆, 李航天, 蔡广研. 不同等级体力活动对慢性肾脏病患者预后的影响[J/OL]. 中华肾病研究电子杂志, 2024, 13(03): 121-128.
[9]	肖伍豪, 刘抗寒. 晚期慢性肾脏病患者骨质疏松症的治疗研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(02): 92-96.
[10]	吴燕升, 张先闻, 王琳. 慢性肾脏病患者肠道微生态与免疫的关系研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(02): 101-105.
[11]	张轶男, 朱国贞. 急性肾损伤向慢性肾脏病转变研究进展[J/OL]. 中华肾病研究电子杂志, 2024, 13(02): 106-112.
[12]	奚培培, 周加军. 慢性肾脏病患者肌少症机制和诊治的研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(05): 491-495.
[13]	韦美菊, 潘玲. 肠道菌群-胆汁酸代谢轴在慢性肾脏病中的研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(02): 219-222.
[14]	袁蔡骏, 闻萍, 徐玲玲. 连续血糖监测在慢性肾脏病合并糖尿病患者中的应用研究进展[J/OL]. 中华临床医师杂志(电子版), 2024, 18(01): 79-82.
[15]	罗婷, 邱令智, 易东, 鄢华. 线粒体功能障碍与心血管疾病、缺血性脑卒中及慢性肾脏病关系的研究进展[J/OL]. 中华脑血管病杂志(电子版), 2024, 18(01): 60-63.

阅读次数

全文

摘要

选择文件类型/文献管理软件名称

选择包含的内容

Analysis of tryptophan-kynurenine pathway in 47 non-dialysis patients with chronic kidney disease by liquid chromatography-mass spectrometry

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

Analysis of tryptophan-kynurenine pathway in 47 non-dialysis patients with chronic kidney disease by liquid chromatography-mass spectrometry