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二维纳米液相色谱-串联质谱法对针刺治疗绝经后干眼症患者泪液的蛋白质组学分析

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

作者信息

Liu Qingyu, Liu Junling, Ren Chengda, Cai Wenting, Wei Qingquan, Song Yi, Yu Jing

机构信息

Department of Ophthalmology, Shanghai Tenth People's Hospital, Tongji University School of Medicine, Shanghai.

Department of Ophthalmology, Nanchang University, Nanchang.

出版信息

Int J Nanomedicine. 2017 Feb 28;12:1663-1671. doi: 10.2147/IJN.S126968. eCollection 2017.

Abstract

BACKGROUND

The purpose of this study was to investigate whether acupuncture is effective at treating dry eye disease among postmenopausal women and to identify the possible mechanisms.

METHODS

Twenty-eight postmenopausal women with dry eye disease were randomly divided into two groups: an acupuncture plus artificial tears (AC + AT) group and an artificial tears (AT) only group. After baseline examination of clinical parameters and tear sample collection, each patient received the designated modality of topical therapy for 2 months. Post-treatment documentation of clinical parameters was recorded, and tear samples were collected. Tear samples from the AC + AT group were subjected to two-dimensional nano-liquid chromatography coupled with tandem mass spectrometry (2D nano-LC-MS/MS). Western blot analysis was also performed on tear samples from both groups.

RESULTS

After treatment, the Ocular Surface Disease Index scores, symptom assessment scores, scores of sign assessment, and tear break-up time were significantly improved in both groups (=0.000). Symptom assessment scores were significantly improved in the AC + AT group (=0.000) compared with the AT group. 2D nano-LC-MS/MS identified 2,411 proteins, among which 142 were downregulated and 169 were upregulated. After combined AC + AT treatment, the abundance of secreted proteins was increased, whereas that of cytoplasmic proteins decreased (Pearson's test, =0.000, =0.000, respectively). Proteins involved in immunity and regulation were also more abundant (Pearson's test, =0.040, =0.016, respectively), while components and proliferation-related proteins were downregulated (Pearson's test, =0.003, =0.011, respectively).

CONCLUSION

AC + AT treatment increased protein synthesis and secretion, and improved clinical symptoms. These results indicate that acupuncture may be a complimentary therapy for treating postmenopausal dry eye disease.

摘要

背景

本研究旨在调查针灸治疗绝经后女性干眼症是否有效,并确定其可能的作用机制。

方法

28名患有干眼症的绝经后女性被随机分为两组:针灸联合人工泪液(AC + AT)组和单纯人工泪液(AT)组。在对临床参数进行基线检查并采集泪液样本后,每位患者接受指定的局部治疗方式,为期2个月。记录治疗后的临床参数,并采集泪液样本。AC + AT组的泪液样本进行二维纳米液相色谱 - 串联质谱分析(2D nano-LC-MS/MS)。对两组的泪液样本也进行了蛋白质免疫印迹分析。

结果

治疗后,两组的眼表疾病指数评分、症状评估评分、体征评估评分和泪膜破裂时间均显著改善(P = 0.000)。与AT组相比,AC + AT组的症状评估评分显著改善(P = 0.000)。2D nano-LC-MS/MS鉴定出2411种蛋白质,其中142种下调,169种上调。AC + AT联合治疗后,分泌蛋白的丰度增加,而细胞质蛋白的丰度降低(Pearson检验,P分别为0.000和0.000)。参与免疫和调节的蛋白质也更丰富(Pearson检验,P分别为0.040和0.016),而成分和增殖相关蛋白下调(Pearson检验,P分别为0.003和0.011)。

结论

AC + AT治疗增加了蛋白质的合成和分泌,并改善了临床症状。这些结果表明,针灸可能是治疗绝经后干眼症的一种辅助疗法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/12f2/5339014/7744836901da/ijn-12-1663Fig1.jpg

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