LAMA1基因与单纯性高度近视的相关性研究

The correlation between the LAMA1 gene and pathological myopia

目的分析单纯性高度近视眼及非近视眼巩膜组织中的LAMA1基因在转录水平(mRNA水平)是否存在显著性差异,探讨其与病理性近视的相关性。方法收集8例行抗青光眼手术的原发性开角性青光眼患者(伴有单纯性高度近视)、5例自愿捐献角膜的健康正常人(正视眼)及5例行抗青光眼手术的原发性开角性青光眼患者(不伴有单纯性高度近视)术中切除的眼前节巩膜组织。提取所收集的巩膜组织中总RNA,针对LAMA1基因设计特异性引物进行RT-PCR检查(reverse transcription-polymerase chain reaction),用凝胶分析系统测定各样本β-actin和LAMA1基因PCR反应产物电泳条带灰度值,以β-actin电泳条带为参照,求出各样本LAMA1基因PCR反应产物电泳条带灰度值和β-actin电泳条带灰度值的比值(LAMA1/β-actin),继而对所得比值采用单因素方差分析(one-wayANOVA)等统计学方法进行统计分析。结果各样本LAMA1基因PCR反应产物电泳条带灰度值和β-actin电泳条带灰度值的比值(LAMA1/β-actin),在正常对照组[(1048.36±31.75)×10-4]和青光眼组[(1058.29±45.62)×10-4]之间比较差异无显著性(P>0.05),而在单纯性高度近视组[(20.50±21.74)×10-4]明显低于非近视组(正常对照组和青光眼组),两组差异有显著性(P<0.01)。结论病理性近视候选基因的LAMA1基因,在单纯性高度近视眼的巩膜组织中其转录水平明显降低,初步判定LAMA1基因与病理性近视具有相关性。但其在转录水平受抑制的机制及如何导致病理性近视的形成有待进一步研究。

Objective To investigate the correlation between the LAMA1 gene and pathological myopia by analyzing the transcriptional level （mRNA level） of the LAMA1 gene in scleral tissue. Methods Front scleral tissues were used as samples for analysis. Of all the samples used, 8 samples were from eyes of primary open-angle glaucoma （POAG） patients （with simple high myopia） who accepted trabeculectomy; 5 samples were from healthy volunteers with emmetropia who donated their corneas; 5 samples were from POAG patients without high myopia who accepted trabeculectomy. The samples were composed of laminal scleral tissue obtained during trabeculectomy. After extracting the total RNA from each sample, specific primers directly targeting the LAMA1 gene were designed. RT-PCR （reverse transcription-polymerase chain reaction） the gel analytical system was used to measure the electrophoresis strips＇ gray scale values of the PCR reaction products of β-actin and the LAMA1 gene. The ratio of the electrophoresis strips＇ gray scale values of the PCR reaction products of β-actin over the electrophoresis strips＇ gray scale values of PCR reaction products of the LAMA1 gene and β-actin （LAMA1/β-actin） was obtained for ANOVA analysis. Results LAMA1/β-actin of POAG eyes with simple high myopia was significantly lower than that of normal eyes and POAG eyes （P〈0.01） and there was no significant difference between normal eye and POAG eyes （P 〉0.05）. Conclusion The mRNA level of the LAMA1 gene, which is a confirmed candidate gene in pathological myopia, is lower in pathological myopic scleral tissue than in non-myopic scleral tissue. Thus, there was a rela- tionship between the LAMA1 gene and simple high myopia. But the mechanism of the LAMA1 gene, which is suppressed at the transcription level, is not very clear and further pathogenic research should be performed.