Abstract：
BACKGROUND: Corneal tissues indirectly obtain nutritional needs and oxygen to maintain their homeostasis, and therefore, benzalkonium chloride (BAC) containing ocular instillations for medical therapy may, in turn, induce toxic effects more than expected in corneal tissues, especially the inside stroma layer.
METHODS: To evaluate the effects of very low concentrations (10-8%, 10-6%, or 10-4%) of BAC on human corneal stroma, we used two-dimensional (2D) cultures of human corneal stromal fibroblast (HCSF) cells and carried out the following analyses: (1) cell viability measurements, (2) Seahorse cellular bio-metabolism analysis, and (3) the expression of ECM molecules and endoplasmic reticulum (ER) stress-related molecules.
RESULTS: In the absence and presence of 10-8%, 10-6%, or 10-4% concentrations of BAC, cell viability deteriorated and this deterioration was dose-dependent. The results showed that maximal mitochondrial respiration was decreased, the mRNA expression of most of ECM proteins was decreased, and ER stress-related molecules were substantially and dose-dependently down-regulated in HCSFs by the BAC treatment.
CONCLUSIONS: The findings reported herein indicate that the presence of BAC, even at such low concentrations, is capable of causing the deterioration of cellular metabolic functions and negatively affecting the response to ER stress in HCSF cells resulting in a substantially decreased cellular viability.
METHODS: To evaluate the effects of very low concentrations (10-8%, 10-6%, or 10-4%) of BAC on human corneal stroma, we used two-dimensional (2D) cultures of human corneal stromal fibroblast (HCSF) cells and carried out the following analyses: (1) cell viability measurements, (2) Seahorse cellular bio-metabolism analysis, and (3) the expression of ECM molecules and endoplasmic reticulum (ER) stress-related molecules.
RESULTS: In the absence and presence of 10-8%, 10-6%, or 10-4% concentrations of BAC, cell viability deteriorated and this deterioration was dose-dependent. The results showed that maximal mitochondrial respiration was decreased, the mRNA expression of most of ECM proteins was decreased, and ER stress-related molecules were substantially and dose-dependently down-regulated in HCSFs by the BAC treatment.
CONCLUSIONS: The findings reported herein indicate that the presence of BAC, even at such low concentrations, is capable of causing the deterioration of cellular metabolic functions and negatively affecting the response to ER stress in HCSF cells resulting in a substantially decreased cellular viability.
摘要：
背景：角膜组织间接获得营养需求和氧气以维持其稳态，因此,苯扎氯铵（BAC）含有用于药物治疗的眼部滴注剂，反过来,在角膜组织中引起比预期更多的毒性作用，尤其是内部基质层。
方法：为了评估非常低的浓度（10-8％，10-6%,或10-4％）人角膜基质上的BAC，我们使用人角膜基质成纤维细胞（HCSF）细胞的二维（2D）培养，并进行了以下分析：（1）细胞活力测量，(2)海马细胞生物代谢分析,(3)ECM分子和内质网(ER)应激相关分子的表达。
结果：在不存在和存在10-8%的情况下，10-6%,或10-4%浓度的BAC，细胞活力恶化，这种恶化是剂量依赖性的。结果表明，线粒体最大呼吸减少，大多数ECM蛋白的mRNA表达降低，和ER应激相关分子在HCSFs中通过BAC处理基本上和剂量依赖性地下调。
结论：本文报道的研究结果表明，BAC的存在，即使在如此低的浓度下，能够引起细胞代谢功能的恶化并负面影响HCSF细胞对ER应激的反应,导致细胞活力显著降低。
方法：为了评估非常低的浓度（10-8％，10-6%,或10-4％）人角膜基质上的BAC，我们使用人角膜基质成纤维细胞（HCSF）细胞的二维（2D）培养，并进行了以下分析：（1）细胞活力测量，(2)海马细胞生物代谢分析,(3)ECM分子和内质网(ER)应激相关分子的表达。
结果：在不存在和存在10-8%的情况下，10-6%,或10-4%浓度的BAC，细胞活力恶化，这种恶化是剂量依赖性的。结果表明，线粒体最大呼吸减少，大多数ECM蛋白的mRNA表达降低，和ER应激相关分子在HCSFs中通过BAC处理基本上和剂量依赖性地下调。
结论：本文报道的研究结果表明，BAC的存在，即使在如此低的浓度下，能够引起细胞代谢功能的恶化并负面影响HCSF细胞对ER应激的反应,导致细胞活力显著降低。
