受体相互作用蛋白激酶1在类风湿关节炎发病中的作用及其遗传易感性

李心亚; 刘亚玲; 唐玉洁; 李晓玲; 陶金辉

doi:10.16462/j.cnki.zhjbkz.2021.08.013

受体相互作用蛋白激酶1在类风湿关节炎发病中的作用及其遗传易感性

doi: 10.16462/j.cnki.zhjbkz.2021.08.013

230001 合肥，中国科学技术大学附属第一医院(安徽省立医院)风湿免疫科

基金项目:

国家自然科学基金 81771774

安徽省重点研究与开发计划项目 201904a07020103

详细信息

通讯作者:
陶金辉，E-mail: taojinhui@ustc.edu.cn

中图分类号: R593.22
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出版历程
- 收稿日期: 2020-12-30
- 修回日期: 2021-04-02
- 网络出版日期: 2021-08-24
- 刊出日期: 2021-08-10

Role of RIPK1 in the pathogenesis of rheumatoid arthritis and analysis of its geneticsusceptibility

Department of Rheumatology and Immunology, The First Affiliated Hospital of University of Science and Technology of China (Anhui Provincial Hospital), Hefei 230001, China

Funds:

National Natural Science Foundation of China 81771774

The Anhui Key Research and Development Foundation 201904a07020103

More Information

Corresponding author: TAO Jin-hui, E-mail: taojinhui@ustc.edu.cn

摘要

摘要: 目的分析受体相互作用蛋白激酶1(receptor interacting serine/threonine protein kinase 1, RIPK1)在类风湿关节炎(rheumatoid arthritis, RA)发病中的作用及其基因多态性与RA遗传易感性的关联。方法采用实时定量PCR(quantitative real-time PCR, qRT-PCR)方法检测RA和健康对照者的外周血单个核细胞(peripheral blood mononuclear cell, PBMC)中RIPK1的mRNA水平，分析其在RA发病中的作用。在RIPK1调控区和蛋白修饰功能区中筛选单核苷酸多态性(single nucleotide polymorphism, SNP)位点，筛选了rs111393904、rs115041708、rs1158343326、rs116040763、rs116603010、rs116696494、rs1186968649、rs1189800403、rs1193251671、rs1255389565、rs138277932、rs17548629、rs200610530、rs547953577、rs557588787、rs561972571、rs760405932、rs77736895和rs934053809共19个位点，采用iMLDR技术进行基因分型，分析RIPK1基因多态性与RA遗传易感性的关联。结果比较88例RA患者和85例健康对照者RIPK1 mRNA表达水平，结果显示RA组低于健康对照组(Z=3.152, P < 0.000 1)；进一步比较RA活动组(50例)、RA稳定组(38例)和健康对照组三组间差异，结果显示RA活动组RIPK1 mRNA表达水平低于稳定组(Z=2.987, P=0.008 4)和健康对照组(Z=4.247, P < 0.000 1)，RA稳定组与健康对照组间差异无统计学意义(Z=0.584, P>0.999 0)。对103例RA患者和90例健康对照者的SNP位点进行基因分型，结果显示RA患者上述SNP位点基因型和等位基因频率与健康对照组的差异均无统计学意义(均有P>0.05)。结论 RIPK1对RA病情活动可能有保护作用，但其基因多态性与RA遗传易感性可能无关联。
- 类风湿关节炎 /
- 受体相互作用蛋白激酶1 /
- 单核苷酸多态性
Abstract: Objective To analyze the role of receptor interacting protein kinase 1 (RIPK1) in the pathogenesis of rheumatoid arthritis (RA) and exploring the association between gene polymorphism and genetic susceptibility to RA. Methods The mRNA level of RIPK1 in peripheral blood mononuclear cells (PBMC) of RA and healthy controls was detected by quantitative real-time PCR (qRT-PCR), and its role in the pathogenesis of RA was analyzed. Single nucleotide polymorphism (SNP) sites were screened in RIPK1 regulatory region and protein modification functional region, which include rs111393904, rs115041708, rs1158343326, rs116040763, rs11660010, rs116696494, rs1186968649, rs1189800403, rs1193251671, rs1255389565, rs138277932, rs17548629, rs200610530, rs547953577, rs557588787, rs561972571, rs760405932, rs77736895 and rs934053809. The association between RIPK1 gene polymorphism and RA susceptibility was analyzed by iMLDR. Results A total of 88 RA patients and 85 healthy controls were selected, and the expression level of RIPK1 gene in the samples was analyzed. It was found that the expression level of RIPK1 gene in the RA group was lower than that in the healthy control group (Z=3.152, P < 0.000 1). According to the active degree, RA patients were divided into RA active group (50 cases) and RA stable group (38 cases). It was found that the expression level of RIPK1 gene in the RA active group was lower than that in the RA stable group (Z=2.987, P=0.008 4) and healthy control group (Z=4.247, P < 0.000 1). There was no significant difference exist between the RA disease stable group and the healthy control group (Z=0.584, P>0.999 0). A total of 103 RA patients and 90 healthy controls were selected to extract DNA from the PBMC of each group. The genotyping results showed that the difference in genotype and allele frequency of the above SNP loci in RIPK1 gene in RA patients was not statistically significance compared with the healthy controls (all P>0.05). Conclusion RIPK1 may have a protective effect on RA disease activity, but its genetic polymorphism may not be associated with RA genetic susceptibility.
- Rheumatoid arthritis /
- Receptor interacting serine/threonine protein kinase 1 /
- Single nucleotide polymorphism

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图 1 RA活动组、RA稳定组与HC组RIPK1的mRNA表达水平比较

注：^a P < 0.01，^b P < 0.0001。

Figure 1. Comparison of RIPK1 mRNA expression among RA active group, RA stable group and HC group

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表 1 RIPK1翻译后修饰位点对应的SNP位点

Table 1. SNP sites corresponding to post-translation modification sites of RIPK1

氨基酸位点	功能	SNP
S14^[10]	激酶活化标志	rs561972571
S15^[10]	激酶活化标志	rs1186968649
K45^[2]	突变失去激酶活性	rs111393904
S166^[10]	激酶活化标志	rs138277932
T189^[2]	抑制激酶激活	rs547953577，rs1193251671
S330^[10]	磷酸化	rs1255389565
S333^[5]	磷酸化，抑制激酶激活	rs557588787
S335^[5]	磷酸化，抑制激酶激活	rs1158343326
K377^[5]	激活TAK1，存活，抑制坏死	rs760405932
D138^[10]	突变抑制激酶活性，挽救TBK1缺失的致死	rs1189800403，rs934053809

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表 2 RIPK1关键SNP位点相关信息

Table 2. RIPK1 key SNP sites related information

SNP	染色质	染色质位点	功能变化	等位基因
rs17548629	6	3114457	-	C/T
rs116040763	6	3113491	p.Thr645Met	C/T
rs1193251671	6	3083425	p.Thr189Ile	C/T
rs200610530	6	3105730	p.Glu341Ser	G/A
rs116603010	6	3081259	p.Ile123Thr	T/C
rs1158343326	6	3090980	-	A/G
rs77736895	6	3113880	-	C/T
rs111393904	6	3077192	p.Lys45Asn	A/C
rs760405932	6	3105839	p.Lys377Thr	A/C
rs934053809	6	3081305	p.Asp138Glu	C/G
rs1186968649	6	3077102	p.Ser15Arg	T/G
rs138277932	6	3083357	p.Ser166Arg	C/G
rs116696494	6	3106044	p.His445Gln	T/A
rs115041708	6	3083400	p.Gly181Ser	G/A/T
rs1255389565	6	3104532	p.Ser330Ter	C/A/T
rs1189800403	6	3081303	p.Asp138Tyr	G/T
rs547953577	6	3083424	p.Thr189Pro	A/C
rs561972571	6	3077097	p.Ser14Ala	T/G
rs557588787	6	3090974	-	C/T

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表 3 RA和HC间RIPK1不同SNP位点基因型及等位基因频率分布的差异

Table 3. Allele and genotype frequencies of SNPs in the RIPK1 gene in RA patients

SNP	遗传模型	RA	HC	χ²值	OR(95% CI)值	P值
rs17548629	C vs. T	159/47	134/46	0.394	1.161(0.728~1.853)	0.530
	CC vs. TT	62/6	50/6	0.126	1.240(0.377~4.081)	0.723
	CT vs. TT	35/6	34/6	0.002	1.029(0.302~3.508)	0.963
	CC+CT vs. TT	97/6	84/6	0.058	1.155(0.359~3.716)	0.809
	CC vs. CT+TT	62/41	50/40	0.424	1.210(0.682~2.146)	0.515
rs77736895	C vs. T	198/8	168/12	1.515	1.768(0.706~4.427)	0.218
	CC vs. TT	10/0	79/1	0.126	1.013(0.988~1.038)	0.722
	CT vs. TT	8/0	10/1	0.768	1.100(0.913~1.326)	0.381
	CC+CT vs. TT	103/0	89/1	1.150	1.011(0.989~1.034)	0.283
	CC vs. CT+TT	95/8	79/11	1.074	1.653(0.634~4.311)	0.300
rs116696494	A vs. T	2/204	1/179	0.215	1.755(0.158~19.516)	0.643
	AA vs. TT	0/101	0/89
	AT vs. TT	2/101	1/89	0.217	1.762(0.157~19.766)	0.642
	AA+AT vs. TT	1/101	1/89	0.008	0.881(0.054~14.295)	0.929
	AA vs. AT+TT	0/103	0/90

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