University of Alabama at Birmingham School of Optometry Department of Optometry and Vision Science Birmingham, AL.
University of Alabama at Birmingham School of Medicine Department of Pharmacology and Toxicology Birmingham, AL.
Mol Vis. 2023 Nov 6;29:289-305. eCollection 2023.
The purpose of this study was to explore the effects of a PGF analog, latanoprost, and its preservative, benzalkonium chloride (BAK), on the cell viability and lipidomic expression of immortalized human meibomian gland epithelial cells (HMGECs).
Differentiated HMGECs were exposed to latanoprost (0.05 to 50 µg/ml), BAK (0.2 to 200 µg/ml), or combined latanoprost-BAK (0.05-0.2 to 50-200 µg/ml). EP- and FP-type receptors, the cognate receptors of PGE and PGF, were inhibited, thereby sparing and isolating the function of each receptor to one condition. Cell viability was assessed by ATP quantitation, and lipid extracts were analyzed by ESI-MSMS with a Triple TOF 5600 Mass Spectrometer (SCIEX, Framingham, MA) using SCIEX LipidView 1.3.
Latanoprost and BAK were found to be lethal to HMGECs at the highest concentrations (p < 0.001 for both). The cytotoxicity of latanoprost was mediated through FP- and EP-independent mechanisms. Both latanoprost and BAK significantly modulated the lipidomic expression of several cholesteryl esters (8% and 30%, respectively) and triacylglycerols (10% and 12%, respectively). The combined latanoprost-BAK agent appeared to be no more toxic and to only negligibly alter the lipid profile relative to its individual components.
The use of latanoprost and BAK in glaucoma may alter the viability of the meibomian glands and their lipid expression in vivo. Sublethal concentrations of BAK appear to modulate meibum lipid expression, particularly in relation to sterol biosynthesis. Non-preserved latanoprost had less cytotoxicity at lower doses and fewer lipidomic effects compared to BAK, further strengthening the argument in favor of BAK-free pharmaceutical preparations.
本研究旨在探讨前列腺素 F 类似物拉坦前列素及其防腐剂苯扎氯铵（BAK）对永生化人眼板腺上皮细胞（HMGEC）活力和脂质组表达的影响。
分化的 HMGEC 暴露于拉坦前列素（0.05 至 50μg/ml）、BAK（0.2 至 200μg/ml）或拉坦前列素-BAK（0.05-0.2 至 50-200μg/ml）联合。抑制 EP 和 FP 型受体，即 PGE 和 PGF 的同源受体，从而保留和分离每种受体对一种条件的功能。通过 ATP 定量评估细胞活力，并用 ESI-MSMS 通过三重四极杆 5600 质谱仪（SCIEX，马萨诸塞州弗雷明汉）进行脂质提取物分析，使用 SCIEX LipidView 1.3。
发现拉坦前列素和 BAK 在最高浓度下对 HMGEC 具有致命作用（两者均为 p<0.001）。拉坦前列素的细胞毒性是通过 FP 和 EP 独立机制介导的。拉坦前列素和 BAK 均显著调节几种胆甾醇酯（分别为 8%和 30%）和三酰基甘油（分别为 10%和 12%）的脂质组表达。联合的拉坦前列素-BAK 制剂似乎毒性更小，与单独成分相比，仅轻微改变脂质谱。
青光眼患者使用拉坦前列素和 BAK 可能会改变眼板腺的活力及其体内脂质表达。亚致死浓度的 BAK 似乎调节了类脂的表达，特别是与固醇生物合成有关。与 BAK 相比，低剂量下非防腐剂拉坦前列素的细胞毒性更小，脂质组学影响也更小，这进一步加强了支持不含 BAK 的药物制剂的论点。
