Division of Endocrinology, Department of Internal Medicine, Faculty of Medicine, University of Debrecen, Debrecen, 4032, Nagyerdei krt. 98, Hungary.
Department of Ophthalmology, Faculty of Medicine, University of Debrecen, Debrecen, 4032, Nagyerdei krt. 98, Hungary.
Chem Biol Interact. 2024 Jun 1;396:111045. doi: 10.1016/j.cbi.2024.111045. Epub 2024 May 9.
Orbital connective tissue changes are contributors to the pathogenesis in thyroid eye disease (TED). Activated fibroblasts respond to immune stimuli with proliferation and increased hyaluronan (HA) production. Cyclosporin A (CsA) was reported to be beneficial in the treatment of TED. PDGF isoforms are increased in orbital tissue of TED patients and enhance HA production. We aimed to study the effect of CsA on HA production and hyaluronan synthase (HAS1, 2 and 3) and hyaluronidase (HYAL1 and 2) mRNA expressions in orbital fibroblasts (OFs). Measurements were performed in the presence or absence of CsA (10 μM) in unstimulated or PDGF-BB (10 ng/ml) stimulated OFs. The HA production of TED OFs (n = 7) and NON-TED OFs (n = 6) were measured by ELISA. The levels of mRNA expressions were examined using RT-PCR. The proliferation rate and metabolic activity were measured by BrdU incorporation and MTT assays, respectively. Treatment with CsA resulted in an average 42% decrease in HA production of OFs (p < 0.0001). CsA decreased the expression levels of HAS2, HAS3 and HYAL2 (p = 0.005, p = 0.005 and p = 0.002, respectively.) PDGF-BB increased HA production (p < 0.001) and HAS2 expression (p = 0.004). CsA could reduce the PDGF-BB-stimulated HA production (p < 0.001) and HAS2 expression (p = 0.005) below the untreated level. In addition, CsA treatment caused a decrease in proliferation potential (p = 0.002) and metabolic activity (p < 0.0001). These findings point to the fact that CsA affects HA metabolism via HAS2, HAS3 and HYAL2 inhibition in OFs. In addition to its well characterized immunosuppressant properties, CsA's beneficial effect in TED may be related to its direct inhibitory effect on basal and growth factor stimulated HA production.
眼眶结缔组织变化是甲状腺眼病 (TED) 发病机制的促成因素。活化的成纤维细胞对免疫刺激物的反应是增殖和透明质酸 (HA) 产生增加。环孢素 A (CsA) 已被报道可用于 TED 的治疗。PDGF 同工型在 TED 患者的眼眶组织中增加，并增强 HA 产生。我们旨在研究 CsA 对眼眶成纤维细胞 (OFs) 中 HA 产生以及透明质酸合酶 (HAS1、2 和 3) 和透明质酸酶 (HYAL1 和 2) mRNA 表达的影响。在无刺激或 PDGF-BB(10ng/ml)刺激的 OFs 中，分别在存在或不存在 CsA(10μM)的情况下进行 HA 产生测量。通过 ELISA 测量 TED OFs(n=7)和 NON-TED OFs(n=6)的 HA 产生。使用 RT-PCR 检查 mRNA 表达水平。通过 BrdU 掺入和 MTT 测定分别测量增殖率和代谢活性。CsA 处理导致 OFs 的 HA 产生平均降低 42%(p<0.0001)。CsA 降低 HAS2、HAS3 和 HYAL2 的表达水平(p=0.005、p=0.005 和 p=0.002)。PDGF-BB 增加 HA 产生(p<0.001)和 HAS2 表达(p=0.004)。CsA 可以将 PDGF-BB 刺激的 HA 产生(p<0.001)和 HAS2 表达(p=0.005)降低至未经处理的水平以下。此外，CsA 处理导致增殖潜力下降(p=0.002)和代谢活性下降(p<0.0001)。这些发现表明 CsA 通过抑制 HAS2、HAS3 和 HYAL2 来影响 OFs 中的 HA 代谢。除了其特征明显的免疫抑制特性外，CsA 在 TED 中的有益作用可能与其对基础和生长因子刺激的 HA 产生的直接抑制作用有关。
