Department of Ophthalmology, Koshigaya Hospital, Dokkyo Medical University, 2-1-50 Minamikoshigaya, Koshigaya, Saitama 343-8555, Japan.
Exp Eye Res. 2012 Mar;96(1):48-54. doi: 10.1016/j.exer.2012.01.001. Epub 2012 Jan 20.
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), β-actin (β-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 β-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 β-ACT after supplementation among male or female patients. β-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.
这是一项初步研究，旨在通过研究晶状体中抗氧化酶的 mRNA 表达来探讨抗氧化剂补充对晶状体过氧化状态的影响。本研究中测量了葡萄糖-6-磷酸脱氢酶 (G6PDH)、β-肌动蛋白 (β-ACT) 和 18S rRNA (18S) 的 mRNA 表达水平，因为它们是通过实时逆转录聚合酶链反应 (RT-PCR) 在各种组织中测量 mRNA 水平的常用参考基因。在获得知情同意后，本研究纳入了 13 名双眼白内障程度相同的患者。在白内障手术中，从前囊膜采集一块组织，同时从前囊膜和晶状体上皮细胞 (LEC) 中采集一个样本作为摄入前样本。手术后第一天开始口服抗氧化补充剂 Ocuvite+叶黄素 (®)。6 周后，在对侧眼白内障手术中从前囊膜采集一块组织，同时从前囊膜和 LEC 中采集一个样本作为摄入后样本。从匀浆样本中提取 RNA，并反转录 cDNA 以测量 mRNA 水平。使用 RT-PCR 测量 G6PDH、18S 和 β-ACT 的表达水平。摄入后样本的 G6PDH 和 18S 表达水平明显高于摄入前样本。在摄入前和摄入后样本中均观察到 G6PDH 和 18S 的表达水平之间存在显著正相关。在进行性别特异性分析后，发现女性患者摄入后样本的 G6PDH 和 18S 表达水平明显更高。男性患者摄入后样本的 G6PDH 和 18S 表达水平之间存在显著正相关。在男性或女性患者中，补充后 β-ACT 的基因表达水平没有显著变化。β-ACT 已被验证可用于通过 RT-PCR 测量晶状体抗氧化剂补充的效果。抗氧化剂补充增加了戊糖磷酸循环中的 G6PDH 和核糖体中的 18S rRNA。
