ABSTRACT
This was a preliminary study of the effects of antioxidant supplementation on the peroxidation status of the lens by investigating mRNA expression of anti-oxidative enzymes in the lens. The mRNA expression levels of glucose-6-phosphate dehydrogenase (G6PDH), ß-actin (ß-ACT) and 18S rRNA (18S) were measured in this study because they are common reference genes for measuring mRNA levels by means of a real-time reverse transcription polymerase chain reaction (RT-PCR) in various tissues. Thirteen patients with binocular cataracts of the same grade were included in the study after giving informed consent. A piece of the anterior capsule, along with a sample of lenticular epithelial cells (LECs), was collected as a pre-intake sample during cataract surgery. Ocuvite + Lutein(®), an antioxidant supplement, was administered orally beginning the day after surgery. Six weeks later, a piece of the anterior capsule along with a sample of LECs, was collected as a post-intake sample during cataract surgery of the opposite eye. RNA was purified from the homogenized samples, and cDNA was reverse transcribed to measure mRNA levels. The expression levels of G6PDH, 18S and ß-ACT were measured using RT-PCR. The expression levels of G6PDH and 18S were significantly higher in the post-intake samples than they were in the pre-intake samples. Significant positive correlations between the expression levels of G6PDH and 18S were observed in both the pre- and post-intake samples. Following gender-specific analyses, the expression levels of G6PDH and 18S in the post-intake samples were found to be significantly higher among the female patients. A significant positive correlation between the expression levels of G6PDH and 18S was observed in the post-intake samples from the male patients. There were no significant changes in the gene expression levels of ß-ACT after supplementation among male or female patients. ß-ACT has been verified for use as a reference gene for measuring the effects of antioxidant supplementation in the lens by RT-PCR. Antioxidant supplementation was noted to increase G6PDH in the pentose phosphate cycle and 18S rRNA in the ribosome.
