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The tear proteomics analysis of conjunctivochalasis

Proteomics – Tränenflüssigkeitsanalyse bei Conjunctivochalasis

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Zusammenfassung

HINTERGRUND: Um die Pathogenese der Conjunctivochalasis zu untersuchen wurden Proteine aus der Tränenflüssigkeit mittels Shotgun-Methode analysiert und mit den Proteinen der Tränenflüssigkeit von gesunden Augen verglichen. METHODE: Es wurde Tränenflüssigkeit von 8 normalen Kontrollpatienten und von 8 Patienten mit Conjunctivochalasis gewonnen. Mit Mikrokapillarpipetten wurden 15 Mikroliter Tränenflüssigkeit aus jedem Auge isoliert. Durch folgenden Untersuchungsschritte wurden die Proteine identifiziert: Trypsinverdauung, Separation der Peptidmischung durch Reverse-phase high-pressure liquid chromatograph (RP-HPCL), gefolgt von Elektro-Spray-Ionisations-Massenspektrometrie (ESI-MS/MS). RESULTATE: Bei Patienten mit Conjunctivochalasis konnten 356 Proteine, bei gesunden Patienten 352, identifiziert werden, wobei 119 Proteine in beiden Gruppen ident waren. Die Proteine der Tränenflüssigkeit wurden mit GOA klassifiziert. Damit konnten in den Tränen der Patienten mit Conjunctivochalasis Proteine gefunden werden, die die Apoptose regulieren bzw. daran beteiligt sind und solche die mit einer Entzündung in Verbindung stehen. In der Kontrollgruppe konnten diese Proteine nicht isoliert werden. Bei Conjunctivochalasis wurde auch Defensin nachgewiesen. SCHLUSSFOLGERUNG: Mit der Shotgun-Methode können Proteine isoliert und analysiert werden. Sie stellt ein neues Verfahren zur Identifikation der Proteine der Tränenflüssigkeit bei Augen mit Conjunctivochalasis dar. Die Ergebnisse unseres Tests lassen vermuten, dass Ursachen dieser Erkrankung Zellapoptose und Entzündung sein könnten.

Summary

OBJECTIVE: To analyze tear proteins by using shotgun strategy, and to study the pathogenesis of conjunctivochalasis by comparing tear proteins between conjunctivochalasis and normal cases. METHODS: Tears were obtained from 8 normal controls and 8 conjunctivochalasis cases. Fifteen micro liters of tears were collected by micro capillary tubes from each eye. Shotgun strategy was used for tear protein analysis. Trypsin digestion in-solution, separation of peptide mixture by reverse-phase high-pressure liquid chromatograph (RP-HPLC) followed by electro spray ionization mass spectrometry (ESI-MS/MS) identification and bio-information analysis were used. RESULTS: 356 proteins were identified in conjunctivochalasis patients and 352 proteins in the normal controls, and among them 119 proteins were the same. The tear proteins were classified with GOA, which found some apoptosis regulation proteins and apoptosis related proteins and inflammatory response related proteins in conjunctivochalasis but not in normal controls group. Defensin was also found in conjunctivochalasis. CONCLUSION: Shotgun strategy can separate and analyze tear proteins effectively, which provides a new method for identifying the tear protein component of conjunctivochalasis. The special component in conjunctivochalasis tears show conjunctivochalasis maybe related to cell apoptosis and inflammation.

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Authors and Affiliations

  1. Department of Ophthalmology, The Affiliated Putuo Hospital of Shanghai Chinese Traditional Medicine University, Shanghai, China

    X.-r. Zhang, M.-h. Xiang, Q.-q. Wu, Q.-s. Li, Y. Xu & A.-g. Sun

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  1. X.-r. Zhang
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  2. M.-h. Xiang
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  3. Q.-q. Wu
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  4. Q.-s. Li
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  5. Y. Xu
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  6. A.-g. Sun
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Correspondence to X.-r. Zhang.

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Zhang, Xr., Xiang, Mh., Wu, Qq. et al. The tear proteomics analysis of conjunctivochalasis. Spektrum Augenheilkd. 22, 288–294 (2008). https://doi.org/10.1007/s00717-008-0285-6

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