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Elastin modulation and modification by homocysteine: a key factor in the pathogenesis of Pseudoexfoliation syndrome?
  1. Manohar Rebecca1,2,
  2. Ramakrishnan Gayathri1,
  3. Renganathan Bhuvanasundar1,
  4. Krishnamoorthy Sripriya3,
  5. Balekudaru Shantha3,
  6. Narayanasamy Angayarkanni1
  1. 1 RS Mehta Jain Department of Biochemistry and Cell Biology, KBIRVO block, Vision Research Foundation, Sankara Nethralaya, Chennai, Tamil Nadu, India
  2. 2 Tamil Nadu Dr MGR Medical University, Chennai, Tamil Nadu, India
  3. 3 Smt. Jadhavbai Nathamal Singhvee Glaucoma Services, Medical Research Foundation, Sankara Nethralaya, Chennai, Tamil Nadu, India
  1. Correspondence to Dr Narayanasamy Angayarkanni, Department of Biochemistry and Cell Biology, Vision Research Foundation, Chennai, Tamil Nadu, India; drak{at}snmail.org

Abstract

Background Pseudoexfoliation syndrome (PXF) is an idiopathic, elastogenesis-associated systemic disease characterised by amyloid-like material aggregates in the eye. Elevated plasma and aqueous humour (aqH) homocysteine (Hcy) is reportedly associated with PXF. This study is aimed to probe Hcy-mediated alterations in elastin expression.

Methodology Lens level of Hcy (total Hcy (tHcy)), mRNA expression of Eln, CBS and MTR in lens capsule, protein expression of elastin in aqH were estimated by enzyme immunoassay, quantitative PCR and western blot, respectively in PXF, PXF with glaucoma (PXF-G) cases, in comparison with cataract-alone disease controls. Human lens epithelial cells (hLECs) were exposed to Hcy and homocysteine thiolactone (HCTL) to evaluate elastin expression in vitro. Furthermore, elastin recombinant protein was incubated with Hcy and HCTL to assess secondary and tertiary structural modifications based on circular dichroism spectroscopy, spectrophotometric and SEM studies.

Results The lens tHcy was significantly high in PXF (p=0.02) and PXF-G (p=0.009). Eln expression was elevated in PXF and PXF-G (p=0.0007). Elastin level in aqH was elevated in PXF (p=0.01) and PXF-G (p=0.002). Hcy (200 µM) and HCTL (1 µM) promoted elastin expression at mRNA level by 36-fold (p=0.02) and 10-fold (p=0.05), respectively, and at protein level by nearly two-fold in cultured hLECs. Secondary structure changes in elastin protein caused by Hcy were evident from 34.11% drop in α-helix and 6.17% gain in β-sheet. Fluorescence, spectral assays and SEM analyses showed aggregation and amyloid formation of elastin with homocysteinylation.

Conclusion The study reveals that lens accumulation of Hcy associated with hyperhomocysteinaemia is characteristic of PXF that augments elastin expression. Hcy causes structural changes promoting elastin aggregation, thereby contributing to defective elastin in PXF and PXF-G.

  • glaucoma
  • aqueous humour
  • Homocysteine
  • Elastin
  • Pseudoexfoliation syndrome

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Footnotes

  • Correction notice Please note this article has been corrected since it was published online first. Author affiliations were incorrectly presented.

  • Contributors MR: patient sample analysis/in vitro studies/data analysis/manuscript preparation/discussion. RG: analysis/patient recruitment. RB: CD spectroscopy. SAV: clinical data/provided clinical specimens. SB: clinical data/provided clinical specimens. NA: study design/data analysis/interpretation/manuscript preparation/discussion.

  • Funding The study is funded by Indian Council of Medical Research ICMR 52/12/2011-BMS and Department of Science and Technology INSPIRE fellowship IF150725, Govt. of India.

  • Competing interests None declared.

  • Patient consent Obtained.

  • Ethics approval Ethics sub-committee (Institutional Review Board).

  • Provenance and peer review Not commissioned; externally peer reviewed.

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