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

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.