丝氨酸蛋白酶原 9 在眼眶成纤维细胞格雷夫斯眼病发病机制中的作用。

Role of Proprotein Convertase Subtilisin/Kexin Type 9 in the Pathogenesis of Graves' Orbitopathy in Orbital Fibroblasts.

机构信息

Yonsei University College of Medicine, Seoul, South Korea.

Department of Ophthalmology, Severance Hospital, Institute of Vision Research, Yonsei University College of Medicine, Seoul, South Korea.

出版信息

Front Endocrinol (Lausanne). 2021 Jan 8;11:607144. doi: 10.3389/fendo.2020.607144. eCollection 2020.

DOI:10.3389/fendo.2020.607144

PMID:33488522

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7821242/

Abstract

BACKGROUND

The proprotein convertase subtilisin/kexin type 9 (PCSK9) has been implicated in the pathogenesis of inflammatory diseases. We sought to investigate the role of PCSK9 in the pathogenesis of Graves' orbitopathy (GO) and whether it may be a legitimate target for treatment.

METHODS

The was compared between GO (n=11) and normal subjects (n=7) in orbital tissue explants using quantitative real-time PCR, and in cultured interleukin-1β (IL-1β)-treated fibroblasts using western blot. Western blot was used to identify the effects of PCSK9 inhibition on IL-1β-induced pro-inflammatory cytokines production and signaling molecules expression as well as levels of adipogenic markers and oxidative stress-related proteins. Adipogenic differentiation was identified using Oil Red O staining. The plasma PCSK9 concentrations were compared between patients with GO (n=44) and healthy subjects (n=26) by ELISA.

RESULTS

The transcript level was higher in GO tissues. The depletion of PCSK9 blunted IL-1β-induced expression of intercellular adhesion molecule 1 (ICAM-1), IL-6, IL-8, and cyclooxygenase-2 (COX-2) in GO and non-GO fibroblasts. The levels of activated nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and phosphorylated forms of Akt and p38 were diminished when PCSK9 was suppressed in GO fibroblasts. Decreases in lipid droplets and attenuated levels of peroxisome proliferator-activated receptor gamma (PPARγ), CCAAT/enhancer-binding protein β (C/EBPβ), and leptin as well as hypoxia-inducible factor 1α (HIF-1α), manganese superoxide dismutase (MnSOD), thioredoxin (Trx), and heme oxygenase-1 (HO-1) were noted when PCSK9 was suppressed during adipocyte differentiation. The plasma PCSK9 level was significantly higher in GO patients and correlated with level of thyrotropin binding inhibitory immunoglobulin (TBII) and the clinical activity score (CAS).

CONCLUSIONS

PCSK9 plays a significant role in GO. The PCSK9 inhibition attenuated the pro-inflammatory cytokines production, oxidative stress, and fibroblast differentiation into adipocytes. PCSK9 may serve as a therapeutic target and biomarker for GO.

摘要

背景

前蛋白转化酶枯草溶菌素/ kexin 9 型（PCSK9）与炎症性疾病的发病机制有关。我们试图研究 PCSK9 在格雷夫斯眼病（GO）发病机制中的作用，以及它是否可能成为治疗的合理靶点。

方法

采用定量实时 PCR 比较 GO（n=11）和正常对照组（n=7）眼眶组织外植体，采用 Western blot 比较白细胞介素-1β（IL-1β）处理的成纤维细胞。Western blot 用于鉴定 PCSK9 抑制对 IL-1β诱导的促炎细胞因子产生和信号分子表达以及脂肪生成标记物和氧化应激相关蛋白水平的影响。采用油红 O 染色鉴定脂肪生成分化。采用 ELISA 比较 GO 患者（n=44）和健康对照组（n=26）的血浆 PCSK9 浓度。

结果

GO 组织中转录水平较高。GO 和非 GO 成纤维细胞中 PCSK9 耗竭可阻断 IL-1β诱导的细胞间黏附分子 1（ICAM-1）、IL-6、IL-8 和环氧化酶-2（COX-2）的表达。GO 成纤维细胞中 PCSK9 抑制后，激活的核因子 kappa-轻链增强子的 B 细胞（NF-κB）和磷酸化形式的 Akt 和 p38 的水平降低。在脂肪细胞分化过程中抑制 PCSK9 时，脂质滴减少，过氧化物酶体增殖物激活受体γ（PPARγ）、CCAAT/增强子结合蛋白 β（C/EBPβ）和瘦素以及缺氧诱导因子 1α（HIF-1α）、锰超氧化物歧化酶（MnSOD）、硫氧还蛋白（Trx）和血红素加氧酶-1（HO-1）的水平降低。GO 患者的血浆 PCSK9 水平明显升高，与促甲状腺激素结合抑制免疫球蛋白（TBII）和临床活动评分（CAS）水平相关。