PLA2R1相关膜性肾病患者血浆差异表达多肽的鉴定

doi:10.3969/j.issn.1673-8640.2023.03.004

检验医学 ›› 2023, Vol. 38 ›› Issue (3): 223-229.DOI: 10.3969/j.issn.1673-8640.2023.03.004

PLA2R1相关膜性肾病患者血浆差异表达多肽的鉴定

王亚捷¹, 祖白玲¹, 甘辰欣¹, 武娇祥¹, 殳洁¹, 杭晨¹, 陈明晖¹, 林芙君², 胡志刚³, 黄飚⁴, 盛慧明¹()

1.上海交通大学医学院附属同仁医院检验科，上海 200050
2. 上海交通大学医学院附属新华医院肾脏风湿免疫科，上海 200092
3. 江南大学无锡医学院附属无锡儿童医院，江苏无锡 214023
4.浙江理工大学生命科学与医药学院，浙江杭州 310018

收稿日期:2022-02-28 修回日期:2023-02-15 出版日期:2023-03-28 发布日期:2023-05-24
通讯作者: 盛慧明
作者简介:盛慧明，E-mail：hmsheng@shsmu.edu.cn。
王亚捷，女，1997年生，硕士，主要从事自身免疫性疾病致病机制研究。
基金资助:
国家自然科学基金项目(82070730);国家自然科学基金项目(81672083);上海市长宁区重点专科项目(20191001);同仁重点培育学科(tr2020xk12)

Identification of plasma differential peptids in PLA2R1-related membranous nephropathy patients

WANG Yajie¹, ZU Bailing¹, GAN Chenxin¹, WU Jiaoxiang¹, SHU Jie¹, HANG Chen¹, CHEN Minghui¹, LIN Fujun², HU Zhigang³, HUANG Biao⁴, SHENG Huiming¹()

1. Department of Clinical Laboratory，Tongren Hospital，Shanghai Jiao Tong University School of Medicine，Shanghai 200050，China
2. Department of Renal Rheumatology and Immunology，Xinhua Hospital，Shanghai Jiao Tong University School of Medicine，Shanghai 200092，China
3. Wuxi Children's Hospital Affiliated to Jiangnan University School of Medicine，Wuxi 214023，Jiangsu，China
4. College of Science and Medicine，Zhejiang Sci-tech University，Hangzhou 310018，Zhejiang，China

Received:2022-02-28 Revised:2023-02-15 Online:2023-03-28 Published:2023-05-24
Contact: SHENG Huiming

摘要/Abstract

摘要：

目的探讨M型磷脂酶A2受体（PLA2R1）相关膜性肾病患者血浆多肽谱的变化。方法选取28例确诊为原发性膜性肾病（PMN）且抗PLA2R1抗体阳性的患者（PMN组）和28名健康体检者（正常对照组）。采用纳升级高效液相色谱串联质谱（nano-HPLC-MS/MS）检测所有研究对象血浆多肽，筛选出PMN患者与正常对照者的差异表达多肽。对差异表达多肽进行生物信息学分析。采用液相色谱串联质谱（LC-MS/MS）对差异表达多肽进行验证。采用Spearman相关分析评估PMN患者血浆C4b与抗PLA2R1-IgG抗体的相关性。结果采用nano-HPLC-MS/MS检出903条多肽，有31条在PMN组和正常对照组之间呈差异表达，其中表达下调15条、表达上调16条；有8条来源于C4b。京都基因与基因组数据库（KEGG）通路分析结果显示，差异表达多肽的前体蛋白涉及的通路主要为补体和凝血级联。LC-MS/MS验证结果显示，PMN组血浆C4b水平显著低于正常对照组（P<0.001）。Spearman相关分析结果显示，PMN患者C4b与抗PLA2R1-IgG抗体无相关性（r=0.03，P=0.877）。结论 C4b或可作为PLA2R1相关膜性肾病患者潜在的生物标志物。

关键词: 多肽, 补体, 高效液相色谱串联质谱, 原发性膜性肾病, M型磷脂酶A2受体

Abstract:

Objective To investigate the changes of plasma peptide profiles of M-type phospholipase A2 receptor（PLA2R1） -related membranous nephropathy. Methods Totally，28 patients with primary membranous nephropathy（PMN） and positive anti-PLA2R1 antibody（PMN group） and 28 healthy subjects（healthy control group） were enrolled. The plasma peptides were determined by nano-high performance liquid chromatography tandem mass spectrometry（nano-HPLC-MS/MS），and the differentially expressed peptides between PMN and healthy control groups were screened. The differentially expressed peptides were analyzed bioinformatically，and liquid chromatography tandem mass spectrometry（LC-MS/MS）was used to verify them. Spearman correlation analysis was used to evaluate the correlation between plasma C4b and anti-PLA2R1-IgG antibodies in PMN patients.Results A total of 903 peptides were found，and 31 of them were expressed differentially between PMN and healthy control groups，with 15 being down-regulated and 16 being up-regulated. Totally，8 of them were from C4b. Kyoto Encyclopedia of Genes and Genomes（KEGG） pathway analysis revealed that the complement and coagulation cascade were primarily involved in the precursor proteins of differentially expressed peptides. According to the verification results，the plasma C4b level in PMN group was lower than that in healthy control group（P<0.001）. The results of Spearman correlation analysis showed that plasma C4b in PMN patients had no correlation with anti-PLA2R1-IgG antibodies（r=0.03，P=0.877）. Conclusions C4b could be a promising new biomarker in patients with PLA2R1-related membranous nephropathy.

Key words: Peptides, Complement, High performance liquid chromatography tandem mass spectrometry, Primary membranous nephropathy, M-type phospholipase A2 receptor

中图分类号:

R446.1

王亚捷, 祖白玲, 甘辰欣, 武娇祥, 殳洁, 杭晨, 陈明晖, 林芙君, 胡志刚, 黄飚, 盛慧明. PLA2R1相关膜性肾病患者血浆差异表达多肽的鉴定[J]. 检验医学, 2023, 38(3): 223-229.

WANG Yajie, ZU Bailing, GAN Chenxin, WU Jiaoxiang, SHU Jie, HANG Chen, CHEN Minghui, LIN Fujun, HU Zhigang, HUANG Biao, SHENG Huiming. Identification of plasma differential peptids in PLA2R1-related membranous nephropathy patients[J]. Laboratory Medicine, 2023, 38(3): 223-229.

图/表 6

参考文献 14

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组别	例数	抗PLA2R1-IgG抗体/（RU/mL）	抗PLA2R1-IgG4抗体/（ng/mL）	C3/（g/L）	C4/（g/L）
正常对照组	28	<14.00	<200.00	1.17±0.18	0.26±0.06
PMN组	28	151.70±124.70	10 910.50±4 799.80	1.26±0.23	0.31±0.17
t值		6.317	11.81	1.637	1.643
P值		<0.001	<0.001	0.107	0.106

组别	例数	抗PLA2R1-IgG抗体/（RU/mL）	抗PLA2R1-IgG4抗体/（ng/mL）	C3/（g/L）	C4/（g/L）
正常对照组	28	<14.00	<200.00	1.17±0.18	0.26±0.06
PMN组	28	151.70±124.70	10 910.50±4 799.80	1.26±0.23	0.31±0.17
t值		6.317	11.81	1.637	1.643
P值		<0.001	<0.001	0.107	0.106

基因名称	蛋白质登记号	多肽序列	PI	相对分子质量	fold change	P值
ACTR3B	ENST00000256001	VDCGTG	3.80	550.58	-8.85	0.008 8
C4b	ENST00000647698	QLNNRQIR	12.00	1 041.18	-3.92	0.038 6
FGA	ENST00000403106	FLAEGGGVR	6.00	905.02	-2.67	0.020 0
C4b	ENST00000647698	NGFKSHALQLN	8.76	1 228.37	-2.61	0.016 5
C4b	ENST00000647698	NGFKSHALQLNN	8.76	1 342.48	-2.59	0.047 5
C4b	ENST00000647698	NGFKSHALQLNNR	11.00	1 498.66	-2.13	0.000 6
C4b	ENST00000647698	SHALQLNNR	9.49	1 052.16	-2.01	0.033 5
ALB	ENST00000621628	KFQNALL	8.75	833.00	-1.96	0.011 3
C4b	ENST00000647698	SHALQLNNRQI	9.49	1 293.45	-1.89	0.012 7
SERPINF2	ENST00000324015	MEPLGRQLTSGP	5.75	1 285.48	-1.49	0.030 6
KMT2A	ENST00000389506	SKTSEKK	9.70	806.91	-1.49	0.043 7
C4b	ENST00000647698	TLEIPGN	4.00	742.83	-1.43	0.003 7
IFT52	ENST00000373030	AAGKAVP	8.80	612.73	-1.35	0.042 6
C4b	ENST00000647698	ALQLNNR	9.79	827.94	-1.25	0.000 8
CDH23	ENST00000224721	GKVIAN	8.75	600.72	-1.24	0.038 5

基因名称	蛋白质登记号	多肽序列	PI	相对分子质量	fold change	P值
ACTR3B	ENST00000256001	VDCGTG	3.80	550.58	-8.85	0.008 8
C4b	ENST00000647698	QLNNRQIR	12.00	1 041.18	-3.92	0.038 6
FGA	ENST00000403106	FLAEGGGVR	6.00	905.02	-2.67	0.020 0
C4b	ENST00000647698	NGFKSHALQLN	8.76	1 228.37	-2.61	0.016 5
C4b	ENST00000647698	NGFKSHALQLNN	8.76	1 342.48	-2.59	0.047 5
C4b	ENST00000647698	NGFKSHALQLNNR	11.00	1 498.66	-2.13	0.000 6
C4b	ENST00000647698	SHALQLNNR	9.49	1 052.16	-2.01	0.033 5
ALB	ENST00000621628	KFQNALL	8.75	833.00	-1.96	0.011 3
C4b	ENST00000647698	SHALQLNNRQI	9.49	1 293.45	-1.89	0.012 7
SERPINF2	ENST00000324015	MEPLGRQLTSGP	5.75	1 285.48	-1.49	0.030 6
KMT2A	ENST00000389506	SKTSEKK	9.70	806.91	-1.49	0.043 7
C4b	ENST00000647698	TLEIPGN	4.00	742.83	-1.43	0.003 7
IFT52	ENST00000373030	AAGKAVP	8.80	612.73	-1.35	0.042 6
C4b	ENST00000647698	ALQLNNR	9.79	827.94	-1.25	0.000 8
CDH23	ENST00000224721	GKVIAN	8.75	600.72	-1.24	0.038 5

基因名称	蛋白质登记号	多肽序列	PI	相对分子质量	fold change	P值
ZNF525	ENST00000491101	PPKKLGL	10.02	751.97	1.33	0.046 5
SMIM35	ENST00000636151	ISTLGLILG	5.52	886.10	1.36	0.037 2
C3orf20	ENST00000253697	QRGRGSP	12.00	756.82	1.40	0.012 5
DENND4C	ENST00000434457	TSISGLVP	5.19	772.90	1.42	0.047 6
ABHD15	ENST00000307201	AALALILAVL	5.57	967.26	1.43	0.044 6
HSD17B4	ENST00000509514	RGSFQVTRAAWEHMKKQKYG	10.45	2 408.77	2.97	0.009 1
BAHCC1	ENST00000675386	ALVAATIN	5.57	771.91	3.58	0.004 7
EIF2AK3	ENST00000303236	AYSGK	8.63	524.57	5.00	0.022 3
DONSON	ENST00000432378	KSTELPV	6.00	772.90	6.08	0.027 1
HP	ENST00000565574	LTTAKNL	8.75	759.90	8.21	0.024 2
ALB	ENST00000621628	IAFAQY	5.52	711.82	11.36	0.032 0
USP6	ENST00000574788	SKGRLRLP	12.01	926.13	12.09	0.004 5
SRPK3	ENST00000393786	IGAEYG	4.00	608.65	16.87	0.006 3
AGAP3	ENST00000622464	TTVKVPGKRLP	11.17	1 195.47	20.65	0.001 4
EFHB	ENST00000344838	ITEIKLPV	6.00	912.14	38.14	0.010 5
LPCAT1	ENST00000283415	AGRKPVRK	12.02	911.12	150.84	0.002 6

PLA2R1相关膜性肾病患者血浆差异表达多肽的鉴定

Identification of plasma differential peptids in PLA2R1-related membranous nephropathy patients

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