利坦色林和度洛西汀对体外原发性阿尼西泮和健康人眼睑基质细胞基因表达的影响

IF 2.6 3区医学 Q2 OPHTHALMOLOGY Ophthalmology and Therapy Pub Date : 2024-11-01 Epub Date: 2024-09-21 DOI:10.1007/s40123-024-01032-8

Zhen Li, Nóra Szentmáry, Fabian N Fries, Shweta Suiwal, Ning Chai, Berthold Seitz, Lei Shi, Maryam Amini, Tanja Stachon

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引用次数: 0

摘要

导言：在配对盒 6（PAX6）突变引起的先天性无角膜症中，PAX6 影响着角膜缘上皮细胞（LECs）的迁移和分化，从而在无角膜症相关角膜病中发挥着关键作用。抗抑郁药物利坦色林（ritanserin）和度洛西汀（duloxetine）会影响LECs中PAX6的表达。支持边缘上皮干细胞的边缘基质细胞在边缘干细胞龛中至关重要。本研究探讨了利坦色林和度洛西汀如何在体外影响先天性无角膜症患者和健康患者的原代人类睑缘基质细胞的基因表达：分离先天性无角膜症患者角膜（AN-LSCs）（n = 8）和健康角膜（LSCs）（n = 8）的原代人类角膜缘基质细胞，并在不含低糖血清（LGSF）或含正常葡萄糖血清（NGSC）的培养基中进行培养。用定量 PCR（qPCR）和 Western 印迹法评估 PAX6、FOSL2、TGF-β1、ACTA2A1、LUM、COL1A1、COL5A1、DSG1、FABP5 和 ADH7 的表达：结果：在LGSF-medium的AN-LSCs中，利坦色林增加了PAX6信使RNA（mRNA）（P = 0.007），降低了TGF-β1和FOSL2 mRNA水平（P = 0.005，P = 0.038）。此外，两种疗法都能降低 TGF-β1 蛋白水平（P = 0.02，P = 0.007），使用利坦色林可提高 FABP5 蛋白水平（P = 0.019）。在中度 LGSF 的 LSCs 中，利坦色林和度洛西汀可降低 ACTA2A1 mRNA 水平（P = 0.028；P = 0.031），而利坦色林治疗可提高 FABP5 mRNA 水平（P = 0.003）。此外，使用度洛西汀可降低α-SMA蛋白水平（P = 0.013），提高FABP5蛋白水平（P = 0.029）。在NGSC-medium的LSCs中，利坦色林可提高LUM、FABP5和ADH7 mRNA和蛋白水平（P = 0.025、P = 0.003、P = 0.047、P = 0.024、P = 0.013、P = 0.039）：我们的研究结果证实，抗精神病药物利坦色林和度洛西汀会改变在LGSF-培养基中培养的AN-LSCs的PAX6和TGF-β1基因表达。研究发现，使用不同的培养基，这些药物对LSCs和AN-LSCs的视黄酸信号通路和角质细胞特征标记都有影响。

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Effect of Ritanserin and Duloxetine on the Gene Expression of Primary Aniridia and Healthy Human Limbal Stromal Cells, In Vitro.

Introduction: In congenital aniridia caused by mutations in paired box 6 (PAX6), PAX6 influences the migration and differentiation of limbal epithelial cells (LECs), thereby playing a pivotal role in aniridia-associated keratopathy. The antidepressants ritanserin and duloxetine affect PAX6 expression in LECs. Limbal stromal cells, which support limbal epithelial stem cells, are crucial in the limbal stem cell niche. This study explores how ritanserin and duloxetine influence gene expression in primary human limbal stromal cells from subjects with congenital aniridia and from healthy subjects, in vitro.

Methods: Primary human limbal stromal cells from corneas affected by aniridia (AN-LSCs) (n = 8) and from healthy corneas (LSCs) (n = 8) were isolated and cultured in either low-glucose serum-free (LGSF) or normal-glucose serum-containing (NGSC) media. Cells were treated with 4 µM ritanserin or duloxetine for 24 h. Quantitative PCR (qPCR) and western blot were used to assess the expression of PAX6, FOSL2, TGF-β1, ACTA2A1, LUM, COL1A1, COL5A1, DSG1, FABP5 and ADH7.

Results: In AN-LSCs with LGSF-medium, ritanserin increased PAX6 messenger RNA (mRNA) (p = 0.007) and decreased TGF-β1 and FOSL2 mRNA levels (P = 0.005, P = 0.038). In addition, TGF-β1 protein levels decreased with both treatments (P = 0.02, P = 0.007), and FABP5 protein level increased, using ritanserin (P = 0.019). In LSCs with LGSF-medium, ACTA2A1 mRNA levels decreased using ritanserin and duloxetine (P = 0.028; P = 0.031), while FABP5 mRNA levels increased with ritanserin treatment (P = 0.003). Also, duloxetine use reduced α-SMA protein (P = 0.013) and increased FABP5 protein levels (P = 0.029). In LSCs with NGSC-medium, ritanserin elevated LUM, FABP5 and ADH7 mRNA and protein levels (P = 0.025, P = 0.003, P = 0.047, P = 0.024, P = 0.013, P = 0.039).

Conclusions: The results of our study confirmed that the antipsychotropic drugs ritanserin and duloxetine alter PAX6 and TGF-β1 gene expression in AN-LSCs cultured in LGSF-medium. These drugs were found to have an impact on retinoic acid signaling pathways and keratocyte characteristic markers both in LSCs and AN-LSCs, using different culture media.

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来源期刊

Ophthalmology and Therapy OPHTHALMOLOGY-

CiteScore

4.20

自引率

3.00%

发文量

157

审稿时长

6 weeks

期刊介绍： Aims and Scope Ophthalmology and Therapy is an international, open access, peer-reviewed (single-blind), and rapid publication journal. The scope of the journal is broad and will consider all scientifically sound research from preclinical, clinical (all phases), observational, real-world, and health outcomes research around the use of ophthalmological therapies, devices, and surgical techniques. The journal is of interest to a broad audience of pharmaceutical and healthcare professionals and publishes original research, reviews, case reports/series, trial protocols and short communications such as commentaries and editorials. Ophthalmology and Therapy will consider all scientifically sound research be it positive, confirmatory or negative data. Submissions are welcomed whether they relate to an international and/or a country-specific audience, something that is crucially important when researchers are trying to target more specific patient populations. 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