不同光环境对兔眼球发育影响机制及中药干预节点的研究

The research based on our previous findings that Daylight lamp delay exposure can result in developmental rabbit eye extension shaft, and follow the " daily clock regulates application of yin and Yang " can effectively intervene it. Further to observe the effect of the different spectral environment and light rhythm on immature rabbit eye development, explore the possible pathogenesis of myopia;the find the " daily clock yin-yang square " in different light conditions can interfere with axial growth and its mechanism. Experiments using one month old New Zealand rabbit, : summer group (the longest sunshine time ) and winter ( the most short time of Sunshine Group), were reared in natural light environment;spectrum Group: control group ( natural light environment ), fluorescent lamp and full spectrum light group, artificial light source groups in optical device, according to the natural light change automatic synchronous control illumination; Biological rhythm group: control group ( natural light rhythm ), daylight lamp delay and full spectrum light delay, delay group light source in the automatic light controller, simulation of modern human life rhythm prolong lighting time; Each group have traditional Chinese medicine group, given to " daily clock Yin and Yang Fang " preparation, all groups for 4 months.From dynamic observation of functional morphology, biochemistry, molecular biology and other indicators on each rabbit eye from childhood to adult stages,by multiple comparison, probe into the influences to rabbits eyes with different light spectrum and different light rhythm , to investigate the possible pathogenesis of myopia.

本课题拟在前期发现日光灯延时照射可造成发育期兔眼眼轴延长，及遵循"阴阳钟调节法"应用"日钟阴阳方"可有效的进行干预的研究基础上，进一步观察不同光谱环境及光照节律对幼兔眼球发育的影响，探讨近视的可能发病机制；同时对"日钟阴阳方"在不同光环境下是否可干预眼轴发育及其机制进行研究。实验采用1月龄新西兰幼兔，年节律组：分夏季组（日照最长时进行）和冬季组（日照最短时进行），均饲养在自然光环境中；光谱组：分对照组（自然光环境）、日光灯和全光谱灯组，人工光源组均在光控器下，参照自然光明暗变化自动同步调控照度；日节律组：分对照组（自然光节律）、日光灯延时和全光谱灯延时组，延时组光源在自动光控器下，模拟现代人类生活节律延长照明时间；各组设中药组，给予"日钟阴阳方"制剂，所有组均进行4个月。实验过程中通过对各组兔眼从幼年到成年各阶段形态学、功能、生化、分子生物学等指标的动态观察，多层次、多方位进行比较、分析。

本课题拟在前期发现光照延时照射可造成发育期兔眼眼轴延长，及应用“日钟阴阳方”可有效的进行干预的研究基础上，观察不同光谱环境及光照节律对幼兔眼球发育的影响，同时对“日钟阴阳方”在不同光环境下是否可干预眼轴发育及其机制进行研究。研究采用1月龄新西兰幼兔，年节律组：分夏季组（日照最长时进行）和冬季组（日照最短时进行），均饲养在自然光环境中；光谱组：分对照组（自然光环境）、LED灯(自然白）和全光谱灯组，人工光源组均在光控器下，参照自然光明暗变化自动同步调控照度；日节律组：分对照组（自然光节律）、LED灯(自然白）延时和全光谱灯延时组，延时组光源在自动光控器下，模拟现代人类生活节律延长照明时间；各组设中药组，给予“日钟阴阳方”制剂，观察时间为3个月。观察了各组幼兔眼球发育、眼组织结构、视功能的影响，如眼轴长度、视网膜电镜等的变化。通过高效液相色谱法检测视网膜多巴胺含量，采用RT-PCR、Western blot、流式细胞技术来检测视网膜钟基因（Cry）及蛋白的表达，及观察各组间观察形态学、功能学、生化学、分子生物学技术的差异。实验结果表明：在人工光环境下（光谱异常、节律异常）的幼兔眼轴较自然光环境下的幼兔眼轴延长，血清多巴胺及视网膜多巴胺减少，差异具有统计学意义，电镜结果表明人工光环境下的幼兔视网膜结构较自然光环境下幼兔差；在阴阳日钟理论指导下的日钟阴阳方可在一定程度上减轻兔眼轴延长，增加血清及视网膜多巴胺含量，并且对视网膜结构有一定的保护作用。