抑制前蛋白转化酶枯草溶菌素6（PCSK6）可能在类风湿关节炎的发展中起保护作用。

Inhibition of PCSK6 may play a protective role in the development of rheumatoid arthritis.

作者信息

Wang Feifei, Wang Lin, Jiang Huiyu, Chang Xiaotian, Pan Jihong

机构信息

From the Shandong Medicinal Biotechnology Centre; the Key Lab for Biotechnology Drugs of Ministry of Health; the Key Lab of Rare and Uncommon Disease; and the Shandong Qianfoshan Hospital, Jinan, China.F. Wang, Masters student; L. Wang, PhD; H. Jiang, Masters student; J. Pan, PhD, Assistant Professor, Shandong Medicinal Biotechnology Centre, and the Key Lab for Biotechnology Drugs of Ministry of Health, and the Key Lab of Rare and Uncommon Disease, Shandong Province; X. Chang, Professor, Shandong Qianfoshan Hospital.

出版信息

J Rheumatol. 2015 Feb;42(2):161-9. doi: 10.3899/jrheum.140435. Epub 2014 Nov 29.

DOI:10.3899/jrheum.140435

PMID:25433529

Abstract

OBJECTIVE

To assess the effect of proprotein convertase subtilisin/kexin type 6 (PCSK6) in the synovial fibroblasts of rheumatoid arthritis (RA). PCSK6 is a proteinase implicated in the proteolytic activity of various precursor proteins and involved in the regulation of protein maturation.

METHODS

PCSK6 expression was detected in the synovial tissue of 10 patients with RA, 10 controls with osteoarthritis, and 10 controls with ankylosing spondylitis using Western blotting and quantitative real-time PCR. Genotyping of 67 tag single-nucleotide polymorphisms (SNP) was performed using an Illumina VeraCode (Illumina) microarray in a case-control study including 267 patients with RA and 160 healthy controls. Genotyping of 4 other tag SNP was performed using a TaqMan probe genotyping assay in 1056 healthy controls and 1151 patients with RA. Cultured RA synovial fibroblasts (RASF) were transfected with PCSK6 small interfering RNA to study changes in the proliferation, invasion, migration capacity, secretion of inflammatory cytokines, cell cycle, and expression profiles of the RASF.

RESULTS

Expression of PCSK6 mRNA and protein was significantly higher in the synovial tissues of individuals with RA than in control tissues. One SNP, rs8029797, was significantly associated with RA (p = 0.011). Knockdown of PCSK6 by RNA interference significantly decreased proliferation, invasion, and migration of RASF. These changes in RASF appeared to be related to reduced tumor necrosis factor-α secretion, G0/G1 arrest, and altered expression of various proteins including those involved in angiogenesis (matrix metalloproteinase 9, nitric oxide synthase trafficking), hypoxia (hypoxia-inducible factor-α, thioredoxin domain containing 5), proliferation (chromosome 10 open reading frame 116), and inflammation [CCL7, chemokine (C-X-C motif) ligand 9, interleukin 26].

CONCLUSION

PCSK6 is upregulated in the synovial tissues of patients with RA and has a genetic effect on the risk of RA. Inhibition of PCSK6 may play a protective role in the development of RA.

摘要

目的

评估前蛋白转化酶枯草溶菌素/克新蛋白酶6型（PCSK6）在类风湿关节炎（RA）滑膜成纤维细胞中的作用。PCSK6是一种蛋白酶，与多种前体蛋白的蛋白水解活性有关，并参与蛋白质成熟的调节。

方法

采用蛋白质免疫印迹法和定量实时聚合酶链反应，检测10例RA患者、10例骨关节炎对照者和10例强直性脊柱炎对照者滑膜组织中PCSK6的表达。在一项病例对照研究中，对267例RA患者和160例健康对照者，使用Illumina VeraCode（Illumina）微阵列对67个标签单核苷酸多态性（SNP）进行基因分型。对1056例健康对照者和1151例RA患者，使用TaqMan探针基因分型法对另外4个标签SNP进行基因分型。用PCSK6小干扰RNA转染培养的RA滑膜成纤维细胞（RASF），以研究RASF的增殖、侵袭、迁移能力、炎性细胞因子分泌、细胞周期及表达谱的变化。

结果

RA患者滑膜组织中PCSK6 mRNA和蛋白的表达明显高于对照组织。一个SNP，rs8029797，与RA显著相关（p = 0.011）。RNA干扰敲低PCSK6可显著降低RASF的增殖、侵袭和迁移能力。RASF的这些变化似乎与肿瘤坏死因子-α分泌减少、G0/G1期阻滞以及包括参与血管生成（基质金属蛋白酶9、一氧化氮合酶转运）、缺氧（缺氧诱导因子-α、含硫氧还蛋白结构域5）、增殖（10号染色体开放阅读框116）和炎症（CCL7、趋化因子（C-X-C基序）配体9、白细胞介素26）的各种蛋白表达改变有关。