肝片形吸虫提取物在体外抑制成纤维样滑膜细胞，并缓解实验性关节炎。

Fasciola hepatica extract suppresses fibroblast-like synoviocytes in vitro and alleviates experimental arthritis.

机构信息

Laboratório de Doenças Autoimunes, Centro de Pesquisa Experimental, Hospital de Clínicas de Porto Alegre, Porto Alegre, Brazil.

Programa de Pós-Graduação em Medicina: Ciências Médicas, Universidade Federal do Rio Grande do Sul, Porto Alegre, Brazil.

出版信息

Adv Rheumatol. 2022 Nov 12;62(1):43. doi: 10.1186/s42358-022-00275-y.

DOI:10.1186/s42358-022-00275-y

PMID:36371346

Abstract

BACKGROUND

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by synovial inflammation, fibroblast-like synoviocytes (FLS) activation and joint destruction. Fasciola hepatica is a platyhelminth that releases excretory-secretory immunomodulatory products capable of suppressing the Th1 immune response. Despite the effectiveness of available treatments for inducing disease remission, current options are not successful in all patients and may cause side effects. Thus, we evaluated the therapeutic potential of F. hepatica extract on FLS from RA patients and arthritis models.

METHODS

FLS were isolated from synovial fluid of RA patients, cultured, and exposed to F. hepatica extract (60, 80, and 100 µg/ml) for different time points to assess cell viability, adherence, migration and invasion. For in vivo experiments, mice with antigen (AIA) and collagen (CIA) induced arthritis received a 200 µg/dose of F. hepatica extract daily. Statistical analysis was performed by ANOVA and Student's t-test using GraphPad Prism 6.0.

RESULTS

In vitro assays showed that extract decreased FLS cell viability at concentration of 100 µg/ml (83.8% ± 5.0 extract vs. 100.0% ± 0.0 control; p < 0.05), adherence in 20% (92.0 cells ± 5.8 extract vs. 116.3 cells ± 7.9 control; p < 0.05), migratory potential (69.5% ± 17.6 extract vs. 100.0% control; p < 0.05), and cell invasiveness potential through the matrigel (76.0% ± 8.4 extract vs. 100.0% control; p < 0.01). The extract reduced leukocyte migration by 56% (40 × 10 leukocytes/knee ± 19.00) compared to control (90.90 × 10 leukocytes/knee ± 12.90) (p < 0.01) and nociception (6.37 g ± 0.99 extract vs. 3.81 g ± 1.44 control; p < 0.001) in AIA and delayed clinical onset of CIA (11.75 ± 2.96 extract vs. 14.00 ± 2.56 control; p = 0.126).

CONCLUSION

Our results point out a potential immunomodulatory effect of F. hepatica extract in RA models. Therefore, the characterization of promising new immunomodulatory molecules should be pursued, as they can promote the development of new therapies. Trial registration Collection of synovial liquid and in vitro procedures were approved by the Ethics Committee with Certificate of Presentation of Ethical Appreciation in Plataforma Brasil (CAAE: 89044918.8.0000.5327; date of registration: 26/07/2018).

摘要

背景

类风湿关节炎（RA）是一种慢性炎症性疾病，其特征为滑膜炎症、成纤维样滑膜细胞（FLS）活化和关节破坏。肝片形吸虫是一种扁形动物，其释放的排泄分泌免疫调节产物能够抑制 Th1 免疫反应。尽管现有治疗方法在诱导疾病缓解方面非常有效，但并非所有患者都能从中受益，而且这些方法可能会引起副作用。因此，我们评估了肝片形吸虫提取物对 RA 患者和关节炎模型中 FLS 的治疗潜力。

方法

从 RA 患者的滑膜液中分离出 FLS，进行培养，并暴露于肝片形吸虫提取物（60、80 和 100µg/ml）不同时间点，以评估细胞活力、黏附、迁移和侵袭。对于体内实验，用抗原（AIA）和胶原（CIA）诱导关节炎的小鼠每天接受 200µg/剂量的肝片形吸虫提取物。使用 GraphPad Prism 6.0 进行方差分析和学生 t 检验进行统计分析。

结果

体外实验表明，提取物在浓度为 100µg/ml 时降低了 FLS 细胞活力（100µg/ml 提取物组为 83.8%±5.0%，对照组为 100.0%±0.0%；p<0.05）、黏附率（20%提取物组为 92.0 个细胞±5.8 个细胞，对照组为 116.3 个细胞±7.9 个细胞；p<0.05）、迁移潜力（69.5%提取物组±17.6 个细胞，对照组为 100.0%个细胞；p<0.05）和穿过基质胶的细胞侵袭潜力（76.0%提取物组±8.4 个细胞，对照组为 100.0%个细胞；p<0.01）。与对照组（90.90×10 个白细胞/膝关节±12.90）相比，提取物减少了白细胞迁移 56%（40×10 个白细胞/膝关节±19.00）（p<0.01）和疼痛感受（6.37g±0.99 提取物 vs. 3.81g±1.44 对照组；p<0.001），并延迟了 CIA 的临床发病时间（11.75±2.96 提取物 vs. 14.00±2.56 对照组；p=0.126）。