PDF(Pubmed)
Abstract：
The purpose of this study was to explore the effects of a PGF2α 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 PGE2 and PGF2α, 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-MSMSALL 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.
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 PGE2 and PGF2α, 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-MSMSALL 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.
摘要：
■这项研究的目的是探索PGF2α类似物的作用，拉坦前列素,和它的防腐剂，苯扎氯铵（BAK），永生化人睑板腺上皮细胞（HMGECs）的细胞活力和脂质组学表达。
■分化的HMGECs暴露于拉坦前列素（0.05至50µg/ml），BAK（0.2至200µg/ml），或联合拉坦前列素-BAK（0.05-0.2至50-200µg/ml）。EP-和FP-型受体，PGE2和PGF2α的同源受体，被抑制，从而将每个受体的功能保留和隔离到一个条件。通过ATP定量评估细胞活力，和脂质提取物用ESI-MSMSALL用三重TOF5600质谱仪(SCIEX，弗雷明汉,MA）使用SCIEXLipidView1.3。
■发现拉坦前列素和BAK在最高浓度下对HMGEC具有致死性（两者均p<0.001）。拉坦前列素的细胞毒性是通过FP和EP非依赖性机制介导的。拉坦前列素和BAK均显着调节几种胆固醇酯的脂质表达（8％和30％，分别)和三酰基甘油(10%和12%，分别)。组合的拉坦前列素-BAK剂似乎不再具有毒性,并且相对于其单个组分仅可忽略地改变脂质分布。
■在青光眼中使用拉坦前列素和BAK可能会改变睑板腺的活力及其在体内的脂质表达。BAK的亚致死浓度似乎可以调节meibum脂质表达，特别是与甾醇生物合成有关。与BAK相比，未保存的拉坦前列素在较低剂量下的细胞毒性较小，脂质体效应较少，进一步加强支持无BAK药物制剂的论点。
■分化的HMGECs暴露于拉坦前列素（0.05至50µg/ml），BAK（0.2至200µg/ml），或联合拉坦前列素-BAK（0.05-0.2至50-200µg/ml）。EP-和FP-型受体，PGE2和PGF2α的同源受体，被抑制，从而将每个受体的功能保留和隔离到一个条件。通过ATP定量评估细胞活力，和脂质提取物用ESI-MSMSALL用三重TOF5600质谱仪(SCIEX，弗雷明汉,MA）使用SCIEXLipidView1.3。
■发现拉坦前列素和BAK在最高浓度下对HMGEC具有致死性（两者均p<0.001）。拉坦前列素的细胞毒性是通过FP和EP非依赖性机制介导的。拉坦前列素和BAK均显着调节几种胆固醇酯的脂质表达（8％和30％，分别)和三酰基甘油(10%和12%，分别)。组合的拉坦前列素-BAK剂似乎不再具有毒性,并且相对于其单个组分仅可忽略地改变脂质分布。
■在青光眼中使用拉坦前列素和BAK可能会改变睑板腺的活力及其在体内的脂质表达。BAK的亚致死浓度似乎可以调节meibum脂质表达，特别是与甾醇生物合成有关。与BAK相比，未保存的拉坦前列素在较低剂量下的细胞毒性较小，脂质体效应较少，进一步加强支持无BAK药物制剂的论点。
