睑板腺功能障碍患者睑脂磷脂成分的LC—MS／MS分析  

Identification of meibomian phospholipids in meibomian gland dysfunction patients using liquidchromatography/tandem mass spectrometry

作　　者：覃冬菊[1] 刘辉[1] 高黎黎[2] 张鹏[2] 徐建江[3] 

机构地区：[1]上海市浦东医院眼科复旦大学附属浦东医院眼科,201399 [2]上海市浦东医院眼科复旦大学附属浦东医院医学科研与创新中心,201399 [3]复旦大学附属眼耳鼻喉科医院眼科,上海200032

出　　处：《中华眼视光学与视觉科学杂志》2015年第11期662-667,共6页Chinese Journal Of Optometry Ophthalmology And Visual Science

基　　金：上海市浦东新区卫生局资助项目（PW2011A-21）;志谢：感谢上海方达医药技术有限公司高江国研究员对研究的指导!

摘　　要：目的用液相色谱．串联质谱联用技术（LC—MS／MS）检测睑板腺功能障碍（meibomian glanddysfunction，MGD）患者睑板腺分泌物磷脂的3种主要成分卵磷脂（溶血磷脂酰胆碱）、脑磷脂（磷脂酰乙醇胺）、神经鞘磷脂与正常人的区别；并分析其相对浓度水平与MGD临床客观体征检测指标的相关性。方法前瞻性配伍组设计。MGD患者22例（22眼）为MGD组，年龄相匹配的正常人30例（30眼）为对照组。MGD组患者选择症状体征较重的眼作为观察对象。用无菌棉签棒按压睑缘后，用无菌的不锈钢刮匙收集睑缘的睑板腺分泌物。所有样本使用LC—MS／MS定性定量分析MGD组睑脂磷脂的主要成分（卵磷脂、脑磷脂、神经鞘磷脂）与正常对照组的区别。2组之间比较采用独立样本t检验，Pearson相关分析MGD组3种磷脂相对浓度水平与睑板腺分泌物性状评分、泪膜破裂时间（BUT）、角膜荧光素染色评分（CFS）、基础泪液分泌试验（SIT）、裂隙灯显微镜下睑缘评分5项指标的关系。结果①睑板腺分泌物3种主要磷脂成分在MGD组与正常对照组间有明显差异，MGD组的睑脂卵磷脂上调（t=2．45，P〈0．05），脑磷脂、神经鞘磷脂下调（t=16．73、18．59，P〈0．05）。②卵磷脂相对浓度水平与睑板腺分泌物性状评分、CFS、裂隙灯显微镜下睑缘评分呈正相关（r=0．602、0．716、0．710，P〈0．01），与BUT、SIT呈负相关（r=-0．628、-0．477，P〈0．05）。脑磷脂与睑板腺分泌物性状评分、CFS、裂隙灯显微镜下睑缘评分呈负相关（r=-0．565、-0．724、-0．702，P〈0．01），与BUT、SIT呈正相关（r=0．659、0．498，P〈0．05）。神经鞘磷脂相对浓度水平与睑板腺分泌物性状评分、CFS、裂隙灯显微镜下睑缘评分呈负相关（r=-0．597、-0．719、-0．698，P〈0．01）；与BUT、SIT呈正相关（r=0．631、0．468，P〈0．05）。结论MGD患�Objective To evaluate the differences in three kinds of meibomian phospholipid composition in healthy subjects and in patients suffering from meibomian gland dysfunction （MGD）. Methods In this prospective study, 22 MGD patients （MGD group） and 30 age and sex matched healthy individuals （control group） were enrolled. One eye of each subject was included in the study. MGD patients underwent sequential examinations as follows： the property of meibomian secretion, tear film break-up time （BUT）, corneal fluorescein staining （CFS）, Schirmer Ⅰ test and lid margin and ocular surface examinations by slit lamp. Meibomian gland secretions were collected using sterile stainless steel curettes in. the healthy control group and MGD group after gentle massage of the lid margin with sterile cotton swabs. The meibomian phospholipids, mainly lysophosphatidylcholine, phosphatidylethanolamine, and sphingomyelin, were detected using liquid chromatography/tandemmass spectrometry （LC-MS/MS）, and comparisons were made between the MGD group and the healthy control group. The data from the two groups were compared by an independent-samples t test and a Pearson correlation analysis was used to assess the correlation between the relative composition concentrations of the three kinds of phospholipids with the clinical parameters of the objective signs of MGD. Results The relative concentration level significantly higher than that in the healthy controls and sphingomyelin levels were lower in the MGD of lysophosphatidylcholine in the MGD group was （t=-2.45, P〈0.05）, while phosphatidylethanolamine group than in healthy controls （t=16.73, P〈0.01 and t=18.59, P〈0.01, respectively）. The relative concentration of lysophosphatidylcholine was positively correlated with the property of meibomian secretion, CFS, lid margin and ocular surface examination by slit lamp （r=-0.602, 0.716, 0.710, P〈0.01, respectively）; while a negative correlation was found with the BUT and Schirmer I test （r=-0.628, -0.477,

关 键 词：睑板腺功能障碍 睑脂 磷脂 液相色谱一串联质谱 

分 类 号：R777.1[医药卫生—眼科]