Effect of tTG AS-ODN on the expression of fibronectin in bovine trabecular meshwork cells

Bo Chen, Yizhen Hu, Yang Cao

Research output: Contribution to journalArticle

Abstract

Objective: To observe the effeets of antisense oligonucleotides against tissue transglutaminase (tTG) on the expression of fibronectin (FN) in bovine trabecular meshwork cells (BTMCs). Methods: The cells were divided into control group, tTG AS-ODN1 group and tTG AS-ODN2 group. After lipofectemine 2 000 reagent delivery, BTMCs were exposed to 0.3 μmol/L antisense tTG and serum-free DMEM respectively. Twenty-four hours later, immunohistochemistry staining was used to detect the effect of antisense tTG on the expression of FN. Western blot and RT-PCR were used for the determine of FN in total cell extracts. Results: immunohistochemistry staining indicated that the mean gray scales were 210.01 ± 3.59 in AS-ODN1 group, 209.23 ± 3.99 in AS-ODN2 group, showing significant differences in comparison with the control group(149.84 ± 5.42) (F = 613.90, P < 0.01). No statistically significant difference was found between AS-ODiN1 group and AS-ODN2 group (P = 0.92). RT-PCR indicated that the FN/β-actin strap central density relative values were 0.68 ± 0.06 in AS-ODN1 group, 0.71 ± 0.05 in AS-ODN2 group with a statistically significant difference compared with the control group (2.45 ± 0.05) (F = 1 357.05, P < 0.01). However, no significant difference between the group of AS-ODN1 and AS-ODN2 (P = 0.73). Western blot indicate that the mean FN/β-actin gray scales were 2.42 ± 0.05 in AS-ODN1 group, 2.44 ± 0.05 in AS-ODN2 group and 1.40 ± 0.06 in control group with a statistically significant difference (F = 517. 37, P < 0.01). There was no statistically significant difference between AS-ODN 1 group and AS-ODN2 group (P = 0. 92). Conclusion: tTG promote the expression of extracellular matrix in BTMCs. Antisense tTG may be used to treat primary open angle glaucoma at the regulation of gene expression through inhibition of the expression of FN and other extracellular matrix.

Original languageEnglish (US)
Pages (from-to)818-822
Number of pages5
JournalChinese Ophthalmic Research
Volume25
Issue number11
StatePublished - Nov 1 2007
Externally publishedYes

Keywords

  • Antisense olignucleotides
  • Fibronectin
  • Primary open angle glaucoma
  • Trabecular meshwork cell
  • Transfer
  • Transglutaminase

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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