Edwards Genea, Aribindi Katyayini, Guerra Yenifer, Bhattacharya Sanjoy K
Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, FL 33136, USA.
Bascom Palmer Eye Institute, University of Miami School of Medicine, Miami, FL 33136, USA.
Biochimie. 2014 Oct;105:99-109. doi: 10.1016/j.biochi.2014.06.019. Epub 2014 Jul 9.
To identify the sphingolipid and ceramide species and their quantitative differences between normotensive and hypertensive intraocular pressure states in DBA/2J mouse aqueous humor (AH).
Normotensive and hypertensive AH was sampled from mice by paracentesis. Lipid extraction was performed using modifications of the Bligh and Dyer method. Protein concentration was estimated using the Bradford colorimetric assay. Sphingolipids and ceramides were identified and subjected to ratiometric quantification using appropriate class specific lipid standards on a TSQ Quantum Access Max triple quadrupole mass spectrometer.
The comparative profiles of normotensive and hypertensive DBA/2J mouse AH showed several species of sphingomyelin, sphingoid base, sphingoid base-1-phosphate (S1P) and ceramides common between them. A number of unique lipids in each of the above lipid classes were also identified in normotensive AH that were absent in hypertensive AH and vice versa.
A number of sphingolipid and ceramide species were found to be uniquely present in normotensive, but absent in hypertensive AH and vice versa. Further pursuit of these findings is likely to contribute towards expanding our understanding of the molecular changes associated with increased intraocular pressure (IOP) and glaucoma.
鉴定DBA/2J小鼠房水（AH）中正常眼压和高眼压状态下的鞘脂和神经酰胺种类及其定量差异。
通过穿刺从小鼠采集正常眼压和高眼压的房水。采用改良的布利格-戴尔法进行脂质提取。使用考马斯亮蓝比色法估算蛋白质浓度。在TSQ Quantum Access Max三重四极杆质谱仪上，使用适当的类别特异性脂质标准品鉴定鞘脂和神经酰胺，并进行比例定量。
正常眼压和高眼压DBA/2J小鼠房水的比较图谱显示，它们之间存在几种鞘磷脂、鞘氨醇、鞘氨醇-1-磷酸（S1P）和神经酰胺。在正常眼压房水中还鉴定出上述每种脂质类别中的一些独特脂质，而这些脂质在高眼压房水中不存在，反之亦然。
发现许多鞘脂和神经酰胺种类在正常眼压房水中独特存在，但在高眼压房水中不存在，反之亦然。进一步探究这些发现可能有助于扩大我们对与眼压升高（IOP）和青光眼相关的分子变化的理解。
