不同光环境对兔视网膜生物钟基因Cry1、Cry2表达的影响及中药干预作用的研究

发布时间：2018-04-03 17:24

本文选题：光环境　切入点：视网膜　出处：《南京中医药大学》2017年硕士论文

【摘要】：目的:本课题通过动物实验研究,探讨不同光环境对兔视网膜生物钟基因Cry1、Cry2表达的影响。结合王育良教授提出的日钟阴阳理论,观察基于阴阳钟理论上的中药复方"日钟阴阳方"对不同光环境下兔视网膜钟基因Cry1、Cry2表达的干预作用。方法:72只1月龄雌雄兼备的新西兰幼兔,均为清洁动物,随机标号分为两大组,分别为4周组和12周组。两大组又随机分为6组,每组6只,即自然光组、正白光LED灯组、全光谱组、自然光中药组、正白光LED灯中药组和全光谱中药组。自然光组:饲养在正常自然光照射的环境中;正白光LED灯组:饲养在正白光LED灯照射的环境下;全光谱组;饲养在全光谱灯光环境中。各中药组:分别饲养在相应的光环境中,并饲养的同时分别予以"日钟阴阳方"灌胃,补阳方于每日10:00灌入,补阴方于每日22:00灌入。幼兔中药灌胃剂量参照《药理实验学》中人和动物间按体表面积折算的等效剂量比值进行换算,按照70kg成人体重标准,一副中药颗粒以300ml水冲调,每日分两次口服计算,成人兔(1.5g)每次服药7ml,幼兔减半为每次3.5ml针管灌胃。自然光组、正白光LED灯组、全光谱组未予以生理盐水灌胃,正常饲养。测定实验光源的光谱参数。第4周、12周提取相应组兔视网膜钟基因Cry1、Cry2。结果:4周和12周后,RT-PCR基因定量检测基因含量,正白光LED灯组兔视网膜Cry1 mRNA、Cry2 mRNA水平皆明显降低(P0.01),正白光LED灯中药组兔视网膜Cry1 mRNA、Cry2 mRNA含量显著高于正白光LED灯组(P0.01)。而中药对自然光组和全光谱组兔视网膜Cry1 mRNA、Cry2 mRNA含量无影响(P0.05)。免疫印迹法显示正白光LED灯组兔视网膜Cry1蛋白、Cry2蛋白水平皆明显降低(P0.01),正白光LED灯中药组兔视网膜Cry1蛋白、Cry2蛋白含量显著高于正白光LED灯组(P0.01)。而中药对自然光组和全光谱组兔视网膜Cry1蛋白、Cry2蛋白含量无影响(P0.05)。结论:正白光LED灯对视网膜钟基因Cry1、Cry2表达水平具有一定的影响。全光谱灯对钟基因Cry1、Cry2表达的影响与自然光相似。"日钟阴阳方"对正白光LED灯下兔视网膜钟基因Cry1、Cry2的表达有一定的干预作用。
[Abstract]:Aim: to investigate the effects of different light environments on the expression of Cry1 and Cry2 in rabbit retina.Based on the theory of Yin and Yang of Sun Zhong put forward by Professor Wang Yuliang, the effect of Rizhong Yin-yang prescription on the expression of Cry1 and Cry2 in rabbit retina under different light environment was observed.Methods Seventy-two one-month old New Zealand rabbits, both male and female, were randomly divided into two groups: 4-week group and 12-week group.The two groups were randomly divided into 6 groups: natural light group, positive white LED lamp group, full spectrum group, natural light Chinese medicine group, positive white light LED lamp group and full spectrum Chinese medicine group.Natural light group: raised in normal natural light environment; positive white light LED lamp group: raised in positive white light LED lamp environment; full spectrum group; raised in full spectrum light environment.Each traditional Chinese medicine group: feeding in the corresponding light environment, and feeding with "Rizhong Yin and Yang prescription", Buyang prescription at 10:00 daily, nourishing Yin prescription at 22:00 daily.Adult rabbits (1.5 g) were treated with 7 ml each time, and young rabbits were treated with 3.5ml every time.Natural light group, white light LED lamp group and full spectrum group were fed with normal saline.Spectral parameters of experimental light source were measured.The corresponding group of rabbit retina clock gene Cry1 and Cry2 were extracted at the 4th week and 12th week.However, Chinese herbal medicine had no effect on the content of Cry2 mRNA in rabbit retina Cry1 mRNAs in natural light group and full spectrum group.However, Chinese herbal medicine had no effect on the content of Cry1 protein Cry2 in the retina of rabbits in natural light group and full spectrum group.Conclusion: the positive white LED lamp has a certain effect on the expression of Cry1 and Cry2 in retina.The effect of full spectrum lamp on the expression of Cry1 gene Cry2 is similar to that of natural light. "Rizhong Yin-Yang Fang had a certain effect on the expression of Cry1 Cry2 in rabbit retina under positive white LED lamp.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R276.7

【参考文献】