比较局部滴眼的双效抗过敏药物的细胞毒性和抗过敏作用。

Comparison of cytotoxicities and anti-allergic effects of topical ocular dual-action anti-allergic agents.

机构信息

Department of Ophthalmology, School of Medicine, Pusan National University, Mulgumup, Yangsan, 50612, Gyeongnam Province, Republic of South Korea.

Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital, Yangsan, South Korea.

出版信息

BMC Ophthalmol. 2019 Nov 8;19(1):217. doi: 10.1186/s12886-019-1228-5.

DOI:10.1186/s12886-019-1228-5

PMID:31703568

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6839072/

Abstract

BACKGROUND

To investigate the cytotoxicities of the topical ocular dual-action anti-allergic agents (alcaftadine 0.25%, bepotastine besilate 1.5%, and olopatadine HCL 0.1%) on human corneal epithelial cells (HCECs) and their anti-allergic effects on cultured conjunctival epithelial cells.

METHODS

A Methylthiazolyltetrazolium(MTT)-based calorimetric assay was used to assess cytotoxicities using HCECs at concentrations of 10, 20 or 30% for exposure durations of 30 min, 1 h, 2 h, 12 h or 24 h. Cellular morphologies were evaluated by inverted phase-contrast and electron microscopy. Wound widths were measured 2 h, 18 h, or 24 h after confluent HCECs monolayers were scratched. Realtime PCR was used to quantify anti-allergic effects on cultured human conjunctival cells, in which allergic reactions were induced by treating them with Aspergillus antigen.

RESULTS

Cell viabilities decreased in time- and concentration-dependent manners. Cells were detached from dishes and showed microvilli loss, cytoplasmic vacuoles, and nuclear condensation when exposed to antiallergic agents; alcaftadine was found to be least cytotoxic. Alcaftadine treated HCECs monolayers showed the best wound healing followed by bepotastine and olopatadine (p < 0.0001). All agents significantly reduced the gene expressions of allergic cytokines (IL-5, IL-25, eotaxin, thymus and activation-regulated chemokine, and thymic stromal lymphopoietin) and alcaftadine had the greatest effect (p < 0.0001 in all cases).

CONCLUSIONS

Alcaftadine seems to have less side effects and better therapeutic effects than the other two anti-allergic agents tested. It may be more beneficial to use less toxic agents for patients with ocular surface risk factors or presumed symptoms of toxicity.

摘要

背景

研究局部眼部双重作用抗过敏药物（0.25%盐酸奥洛他定、1.5%贝他斯汀、0.1%盐酸奥洛他定）对人角膜上皮细胞（HCEC）的细胞毒性及其对培养的结膜上皮细胞的抗过敏作用。

方法

采用噻唑蓝（MTT）比色法，以浓度为 10%、20%或 30%、暴露时间 30 分钟、1 小时、2 小时、12 小时或 24 小时的方式，使用 HCEC 评估细胞毒性。通过倒置相差显微镜和电子显微镜评估细胞形态。在 HCEC 单层细胞划痕后 2 小时、18 小时或 24 小时测量划痕宽度。采用实时 PCR 定量检测培养的人结膜细胞的抗过敏作用，其中通过用曲霉菌抗原处理诱导过敏反应。

结果

细胞活力呈时间和浓度依赖性下降。当暴露于抗过敏药物时，细胞从培养皿中分离出来，表现出微绒毛丧失、细胞质空泡和核浓缩；发现盐酸奥洛他定的细胞毒性最小。经盐酸奥洛他定处理的 HCEC 单层细胞的愈合效果最好，其次是贝他斯汀和盐酸奥洛他定（p<0.0001）。所有药物均显著降低过敏细胞因子（IL-5、IL-25、嗜酸性粒细胞趋化因子、胸腺激活调节趋化因子和胸腺基质淋巴细胞生成素）的基因表达，且盐酸奥洛他定的作用最大（所有情况下均为 p<0.0001）。