Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida 33136, USA.
Invest Ophthalmol Vis Sci. 2013 Apr 30;54(4):3037-44. doi: 10.1167/iovs.12-10517.
We compared phospholipid (phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol) profiles of control and glaucomatous trabecular meshwork (TM) derived from human donors.
Control TM and most primary open angle glaucoma (POAG) TM were collected from cadaver donors. A select subset of POAG surgical TM samples also were collected for analyses. Lipid extraction was performed using a modification of the Bligh and Dyer method, protein concentrations were determined using the Bradford method, and for select samples confirmed with densitometry of PHAST gels. Lipids were identified and subjected to ratiometric quantification using a TSQ quantum Access Max triple quadrupole mass spectrometer with precursor ion scan (PIS) or neutral ion loss scan (NLS), using appropriate class specific lipid standards.
The comparative profiles of phosphatidylcholine, phosphatidylserine, phosphoethanolamine, and phosphatidylinositol between control and glaucomatous TM showed several species common between them. A number of unique lipids in all four phospholipid classes also were identified in control TM that were absent in glaucoma TM and vice versa.
A number of phospholipids were found to be uniquely present in control but absent in glaucomatous TM and vice versa. Compared to a previous study of control and POAG blood, a number of these phospholipids are absent locally (TM), as well as systemically (in blood).
我们比较了来源于人供体的对照和青光眼性小梁网(TM)的磷脂(磷脂酰胆碱、磷脂酰丝氨酸、磷脂酰乙醇胺和磷脂酰肌醇)谱。
对照 TM 和大多数原发性开角型青光眼(POAG)TM 从尸体供体中收集。POAG 手术 TM 样本的一个选择子集也用于分析。使用 Bligh 和 Dyer 方法的修改版进行脂质提取,使用 Bradford 方法测定蛋白质浓度,并使用 PHAST 凝胶的密度测定法对选定样品进行确认。使用具有前体离子扫描(PIS)或中性离子损耗扫描(NLS)功能的 TSQ quantum Access Max 三重四极杆质谱仪对脂质进行鉴定和相对定量分析,使用适当的类特异性脂质标准品。
对照和青光眼 TM 之间的磷脂酰胆碱、磷脂酰丝氨酸、磷酸乙醇胺和磷脂酰肌醇的比较图谱显示它们之间有几个共同的物种。在对照 TM 中还鉴定出所有四个磷脂类别的许多独特脂质,而在青光眼 TM 中则不存在,反之亦然。
发现许多磷脂在对照 TM 中独特存在而在青光眼 TM 中不存在,反之亦然。与之前对对照和 POAG 血液的研究相比,这些磷脂中有许多在局部(TM)和全身(血液)都不存在。