针刺治疗绝经后期干眼患者的疗效及其泪液蛋白质组学研究  

Proteomic analysis of tears following acupuncture treatment for menopausal dry eye based on two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry

作　　者：沈天怡 刘俊伶[1] 蔡雯婷[1] 危清泉 余咚卉 金惠子[1] 梁秀玮 于靖[1] SHEN Tian-Yi;LIU Jun-Ling;CAI Wen-Ting;WEI Qing-Quan;YU Dong-Hui;JIN Hui-Zi;LIANG Xiu-Wei;YU Jing(From the Department of Ophthalmology,the Tenth People’s Hospital of Tongji University,Shanghai 200072,China)

机构地区：[1]同济大学附属第十人民医院眼科,上海市200072

出　　处：《眼科新进展》2018年第10期959-963,共5页Recent Advances in Ophthalmology

基　　金：国家自然科学基金面上项目(编号:81470648)

摘　　要：目的研究针刺疗法治疗绝经后期干眼患者的有效性及其分子机制。方法选取28例(56眼)绝经后期干眼妇女,随机分成单纯人工泪液(artificial tears,AT)治疗组(AT组) 14例28眼及针刺联合人工泪液(acupuncture plus artificial tears,AC+AT)治疗组(AC+AT组) 14例28眼,每组均治疗2个月,并对治疗前后两组患者眼表疾病指数(ocular surface disease index,OSDI)评分、症状评分、体征评分以及泪膜破裂时间(break-up time,BUT)进行分析。同时采集治疗前后各组泪液样本,采用二维纳米液相色谱耦合串联质谱法(two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry,2D Nano-LC-MS/MS)对AC+AT组治疗前后泪液进行蛋白质组学分析,采用蛋白质印记法比较两组泪液蛋白差异。结果两组治疗后OSDI评分、症状评分、体征评分和BUT,AT组分别为39. 08±18. 68、7. 43±2. 08、11. 46±2. 29、(4. 75±1. 74) s,AC+AT组分别为24. 55±17. 43、4. 68±2. 54、11. 21±2. 96、(4. 57±2. 71) s;与治疗前相比均明显改善,差异均有统计学意义(均为P=0. 000)。与AT组相比,针刺治疗后AC+AT组的症状评分明显改善(P=0. 000)。2D Nano-LC-MS/MS法分析了泪液中2411种蛋白,其中142种表达下调,169种表达上调。与AT组相比,针刺治疗后AC+AT组患者泪液中细胞质蛋白表达下调(P=0. 000),分泌蛋白表达上调(P=0. 000)。在蛋白功能方面,与AT组相比,AC+AT组细胞组分蛋白表达下调(P=0. 000),免疫蛋白表达上调(P=0. 040)。结论 AC+AT治疗增加了泪液蛋白质合成及分泌,改善临床绝经后期干眼患者症状。针刺治疗可能成为治疗该类干眼患者的有效辅助疗法。Objective To investigate whether acupuncture is an effective treatment for dry eye among postmenopausal women and identify the molecular mechanisms of acupuncture.Methods Twenty-eight postmenopausal women with dry eye(fifty-eight eyes)were randomly divided into two groups:artificial tears(AT)group including fourteen patients with twenty-eight eyes and acupuncture plus artificial tears(AC+AT)group including fourteen patients with twenty-eight eyes.Each patient received the topical therapy for 2 months.Pretreatment and posttreatment data of ocular surface disease index(OSDI)scores,symptom assessment scores,sign assessment scores,tear break-up time(TBUT)were analyzed.Meanwhile,tear samples of both groups were collected.Tear samples from AC+AT group were subjected to two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry(2D Nano-LC-MS/MS)before and after treatment.Western blot analysis was performed on tear samples from both groups.Results After treatment,scores of OSDI,symptom assessment,sign assessment and TBUT were significant improved in both groups than before therapy(P=0.000),which represented 39.08±18.68,7.43±2.08,11.46±2.29 and(4.75±1.74)s in AT group,while represented 24.55±17.43,4.68±2.54,11.21±2.96,(4.57±2.71)s in AC+AT group,respectively.Compared with AT group,symptom assessment scores were improved than those in AC+AT group significantly(P=0.000).2D Nano-LC-MS/MS identified 2411 proteins,among which 142 kinds of proteins were down-regulated and 169 were upregulated.When compared with AT group,the expression of cytoplasmic proteins was down-regulated,while the expression of secreted proteins was up-regulated in AC+AT group after treatment(both P=0.000).In the aspect of function,the expression of proteins involved in components was down-regulated,while the expression of immunity protein was upregulated(P=0.040).Conclusion AC+AT treatment increases the synthesis and secretion of proteins,and improves clinical symptoms in postmenopausal dry eye,which indicates that acupunctu

关 键 词：针刺 绝经后期 干眼 蛋白质组学 

分 类 号：R777.1[医药卫生—眼科]