Purpose

The purpose of this study was to compare the cholesteryl ester (CE) profiles expressed from human meibomian gland epithelial cells (HMGECs) in response to rosiglitazone-induced differentiation to that of normal human meibum.

Methods

HMGECs were cultured with rosiglitazone (vehicle control, 20 μM, or 50 μM) and fetal bovine serum (FBS, 2% or 10%) for 2 days or 5 days. Following culture, lipid extracts were processed and analyzed by ESI-MSMS^ALL in positive ion mode. CEs were identified using both LipidView 1.2 and PeakView 2.2 (SCIEX, Framingham, MA) and compared to literature reports of CEs in normal human meibum.

Results

There were 34 CEs with carbon number ranging from 14 to 34 detected from HMGECs. Across all conditions, HMGECs provided a CE profile that was 14.0% saturated, 60.6% monounsaturated, and 25.4% polyunsaturated. Culturing with 50 μM rosiglitazone and 2% FBS for 2 days resulted in the greatest number of upregulated saturated and monounsaturated CEs and downregulated polyunsaturated CEs. Five CEs were identified as being the most responsive to 50 μM rosiglitazone: CE 24:1, CE 28:1, CE 26:1, CE 18:1, and CE 22:1.

Conclusion

Although differences in the CE profile exist between meibum and HMGECs, rosiglitazone promotes upregulation of highly expressed meibum-relevant CEs and shifts the saturation level toward a more meibum-like profile. The use of rosiglitazone as a differentiating agent is recommended in HMGEC research, and analysis by ESI-MSMS^ALL is encouraged to differentiate meibum-relevant CEs from other nonpolar distractors detected by vital stains.

中文翻译：

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罗格列酮处理后人睑板腺上皮细胞产生的胆固醇酯饱和

目的

本研究的目的是比较人睑板腺上皮细胞 (HMGECs) 表达的胆固醇酯 (CE) 谱，以响应罗格列酮诱导的正常人睑脂分化。

方法

HMGEC 与罗格列酮（载体对照，20 μM 或 50 μM）和胎牛血清（FBS，2% 或 10%）一起培养 2 天或 5 天。培养后，脂质提取物在正离子模式下通过 ESI-MSMS ^{ALL进行处理和分析。}使用 LipidView 1.2 和 PeakView 2.2 (SCIEX, Framingham, MA) 鉴定 CE，并与正常人类睑脂中 CE 的文献报告进行比较。

结果

从 HMGEC 中检测到 34 个碳数在 14 到 34 之间的 CE。在所有条件下，HMGEC 提供的 CE 曲线为 14.0% 饱和、60.6% 单不饱和和 25.4% 多不饱和。用 50 μM 罗格列酮和 2% FBS 培养 2 天导致上调的饱和和单不饱和 CE 和下调的多不饱和 CE 数量最多。五个 CE 被确定为对 50 μM 罗格列酮最敏感：CE 24:1、CE 28:1、CE 26:1、CE 18:1 和 CE 22:1。

结论

尽管睑脂和 HMGEC 之间的 CE 曲线存在差异，但罗格列酮促进高度表达的睑脂相关 CE 的上调，并将饱和度水平转向更像睑脂的曲线。在 HMGEC 研究中建议使用罗格列酮作为鉴别剂，并鼓励通过 ESI-MSMS ^ALL进行分析，以区分睑脂相关 CE 与其他由生命染色检测到的非极性干扰物。