本文選題：正視化 + 頻閃��； 參考：《復(fù)旦大學(xué)》2013年博士論文

【摘要】：出生后視覺環(huán)境的改變是誘發(fā)屈光發(fā)育的關(guān)鍵原因之一。迄今為止,任何一種人工照明均無法做到對自然光線的完全模擬,人工照明中的頻閃對屈光發(fā)育的影響如何?本研究旨在探討頻閃光對屈光發(fā)育敏感期豚鼠的眼軸和屈光度的影響及其機理。主要方法是通過自動控制技術(shù)模擬不同頻閃頻率、亮度及節(jié)律的頻閃光,利用頻閃光誘導(dǎo)豚鼠近視模型,檢測屈光度及眼軸長度改變,并觀察離體眼球形狀及后極部超微結(jié)構(gòu)改變；通過視覺電生理的變化評估視網(wǎng)膜傳導(dǎo)功能；通過分子生物學(xué)方法觀察豚鼠視網(wǎng)膜視蛋白表達及鞏膜膠原代謝的變化。主要結(jié)果：持續(xù)暴露于人工照明環(huán)境下特定頻率的頻閃光能刺激豚鼠眼球過度發(fā)育,并誘導(dǎo)近視性屈光改變；不同頻率、亮度及節(jié)律的頻閃光對豚鼠眼球發(fā)育的影響存在差異性；視網(wǎng)膜電圖及超微結(jié)構(gòu)改變表明豚鼠眼球視網(wǎng)膜的感光傳導(dǎo)功能下降,并出現(xiàn)感光細胞形態(tài)異常；頻閃組視蛋白表達增加,同時鞏膜基質(zhì)金屬蛋白酶MMP-2表達增高。本文結(jié)論：頻閃光這種異常光環(huán)境對豚鼠眼正視化存在影響,視覺感知過程中頻閃光參數(shù)改變對視覺傳導(dǎo)功能的影響、導(dǎo)致過度正視化最終導(dǎo)致近視的作用,及其中的機制值得進一步研究。具體分述如下： 第一部分頻閃光頻率、亮度及節(jié)律對豚鼠眼屈光發(fā)育的影響 目的：探討持續(xù)暴露于不同刺激頻率、亮度及節(jié)律的低頻率頻閃光對豚鼠屈光度及眼軸長度的影響以及頻閃光不同參數(shù)間的差異性。 對象與方法：實驗對象為2w齡健康雄性豚鼠,使用頻閃調(diào)光器進行頻閃光刺激。本部分共分頻率、亮度及節(jié)律ABC共三個實驗組,A組72只豚鼠隨機分為5個不同頻率的頻閃組和1個對照組(n=12),頻閃組分別予5Hz、1Hz、0.5Hz、0.25Hz、0.1Hz的頻率等時交替頻閃(明暗周期50%),亮?xí)r光照度為6001ux,暗時照度為0,對照組為持續(xù)6001ux光照組。B組48只2w齡豚鼠隨機分為3個不同亮度頻閃組及1個對照組(n=12),頻閃組均以0.5Hz(1s暗,1s亮)的頻率以0-600lux、0-300lux及300-600lux的亮度等時交替頻閃,對照組以6001ux持續(xù)亮度照明。AB兩個研究組所有實驗豚鼠均用同一時控開關(guān)控制照明時間為正常作息節(jié)律6：00-18：00。C組36只2w齡豚鼠隨機分為2個不同節(jié)律的頻閃組及1個對照組(n=12),頻閃組以0.5Hz頻率0-600lux照明亮度頻閃,頻閃暴露與停止頻閃時間分別為12h/12h(正常作息節(jié)律6：00-18：00)和0.5h/0.5h兩種節(jié)律,對照組以600lux持續(xù)照度照明,光照時間為6：00～18：00,與12h/12h頻閃組相同。以上3個實驗組均使用505nm波長發(fā)光二極管,應(yīng)用照度測量儀測量即時照明亮度,各組實驗豚鼠每2w記錄屈光度、眼軸長度及角膜曲率半徑,以首次測量日記錄為0w,持續(xù)觀察時間為12w。應(yīng)用GRAPHPADPRISM Version5.0軟件進行圖表繪制,SPSS17.0進行統(tǒng)計分析,雙眼間差異行配對t檢驗,每只豚鼠均取左眼參數(shù)進行進一步統(tǒng)計分析,組間差異采用單因素方差分析及多重比較,P0.05為差異有統(tǒng)計學(xué)意義。 結(jié)果：實驗前各組豚鼠生物學(xué)參數(shù)差異均無統(tǒng)計學(xué)意義(P0.05)。雙眼間差異無統(tǒng)計學(xué)意義(P0.05)。隨時間延長,ABC三個研究組的頻閃光組與對照組相比均出現(xiàn)屈光度向近視偏移以及眼軸延長(P0.05),各組角膜曲率半徑在整個實驗過程中均逐漸增加,不同時間點各組間差異無統(tǒng)計學(xué)意義(P0.05)。12w時各組間近視屈光度及眼軸長度經(jīng)統(tǒng)計學(xué)比較,A組里5種不同頻率中0.5Hz頻閃組近視程度及眼軸延長最明顯,B組里不同亮度組中屈光度改變及眼軸延長0-6001ux0-3001ux300-6001ux對照組。C組中不同節(jié)律組屈光度改變及眼軸延長0.5h/0.5h12h/12h對照組。 結(jié)論：(1)長期暴露于特定低頻率的頻閃光能刺激豚鼠眼球過度發(fā)育并誘導(dǎo)軸性近視形成,不同的刺激頻率之間有差異性,0.5Hz頻閃光的影響最為顯著。(2)降低頻閃光的刺激強度或明暗波動幅度有助于減緩屈光發(fā)展程度。(3)頻閃暴露的節(jié)律會對眼球的過度發(fā)育造成影響。正常作息節(jié)律改變導(dǎo)致豚鼠眼球過度發(fā)育的現(xiàn)象提示中樞神經(jīng)調(diào)節(jié)機制可能參與其中。 第二部分0.5Hz頻閃光環(huán)境對豚鼠眼球結(jié)構(gòu)、功能的影響及機制 目的：探討持續(xù)0.5Hz頻閃光暴露對豚鼠視覺傳導(dǎo)功能及視網(wǎng)膜鞏膜超微結(jié)構(gòu)的影響及其相關(guān)機制。 對象與方法：36只2w齡雄性豚鼠隨機分為頻閃組及對照組(n=18),頻閃組予頻閃調(diào)光器以0.5Hz頻率(明暗周期50%,即1s亮、1s暗)等時交替頻閃,實驗前先通過照度測量儀測量籠內(nèi)照明亮度,照度波動0-6001ux；對照組通過控制器旋鈕調(diào)節(jié)籠內(nèi)光源照度為持續(xù)6001ux。2組均使用505nm波長發(fā)光二極管,以上2組均由同一時控開關(guān)控制,控制光照時間為正常晝夜12h輪替,光照時間6：00-18：00。實驗3周后兩組豚鼠中各選取6只豚鼠麻醉后取出眼球分離視網(wǎng)膜及鞏膜組織,Western-blot觀察蛋白水平視網(wǎng)膜視蛋白及后部鞏膜MMP-2表達的差異。其余24只豚鼠分別于實驗前及第12w測量豚鼠的視網(wǎng)膜電圖(F-ERG)及振蕩電位(Ops),實驗12w后所有豚鼠過量麻醉處死后取出眼球,測量離體眼球水平徑、垂直徑、前后徑。病理學(xué)光鏡及電鏡下觀察眼球后極部視網(wǎng)膜及鞏膜超微結(jié)構(gòu),與對照組進行比較。應(yīng)用GRAPHPADPRISM Version5.0軟件進行圖表繪制,SPSS17.0進行統(tǒng)計分析,組間差異采用t檢驗,P0.05為差異有統(tǒng)計學(xué)意義。 結(jié)果：與對照組相比,頻閃組各豚鼠眼球視網(wǎng)膜視蛋白及鞏膜MMP-2表達均增加(P0.05)；第12w時與對照組相比,頻閃組的F-ERG的a波潛伏期延長(P0.05)；a波振幅及b波潛伏期及振幅未見統(tǒng)計學(xué)差異(P0.05), Ops兩組間差異未見統(tǒng)計學(xué)意義(P0.05)；眼球水平徑、垂直徑及前后徑分別較對照組增加0.89±0.3mm,0.69±0.2m及0.96+0.3mm,差異有統(tǒng)計學(xué)意義(P0.05),組織學(xué)觀察：光鏡下頻閃光組眼球鞏膜纖維出現(xiàn)擴張,電鏡下視網(wǎng)膜感光細胞層外段排列紊亂并可見較多變形及脫落的外節(jié)盤膜,對照組無明顯異常。 結(jié)論：1.頻閃光誘導(dǎo)的豚鼠近視模型視蛋白表達增加,提示視錐細胞感受異常光覺信息有可能是眼軸變化的起始環(huán)節(jié)。2.頻閃中鞏膜MMP-2增加,與豚鼠眼軸的延長之間的關(guān)系值得進一步探討。3.持續(xù)頻閃光刺激誘導(dǎo)豚鼠眼球過度發(fā)育,可影響視網(wǎng)膜感光細胞的超微結(jié)構(gòu)與傳導(dǎo)功能。
[Abstract]:The changes in visual environment after birth are one of the key causes of refractive development. To date, any artificial lighting can not complete the complete simulation of natural light. How does the stroboscopic effect of artificial lighting affect the refractive development? This study aims to explore the eye axis and diopter of the frequency flashes in the flexion sensitive guinea pigs. The main method is to simulate frequency flashes of different stroboscopic frequencies, luminance and rhythm by automatic control technology. Using frequency flash to induce myopia model of guinea pig, detect the changes of refractive index and axial length, observe the shape of the eyeball and the ultrastructure of the posterior pole, and evaluate the retinal conduction work through the changes of visual electrophysiology. The changes in the retina protein expression and collagen metabolism in the retina of guinea pigs were observed by molecular biology methods. Main results: the frequency flash that sustained exposure to a specific frequency in the artificial lighting environment could stimulate the overdevelopment of the eyeball of guinea pigs and induce the myopic refractive changes; the frequency of frequency, brightness and rhythm of the guinea pig's eyeballs with different frequency, brightness and rhythm The effects of development were different. The change of electroretinogram and ultrastructure showed that the photosensitive conduction function of the retina of the guinea pig was decreased, and the morphologic abnormalities of the photoreceptor cells appeared; the expression of the stroboscopic protein expression increased and the expression of the scleral matrix metalloproteinase MMP-2 increased. In the process of visual perception, the effect of the change of frequency flashing parameters on the visual conduction function in the visual perception process leads to the effect of myopia, and the mechanism in it should be further studied.
Effects of the first frequency, brightness and rhythm on the refractive development of guinea pigs
Objective: To investigate the effects of low frequency frequency flashes on the refractive index and axial length of guinea pigs and the difference between different parameters of frequency flash.
Subjects and methods: the experimental subjects were 2W aged healthy male guinea pigs, using stroboscopic light to stimulate frequency flash stimulation. The total frequency division rate, brightness and rhythm of ABC were three experimental groups, and 72 guinea pigs in group A were randomly divided into 5 different frequency stroboscopic groups and 1 control groups (n=12). The frequency group was given the frequency of 5Hz, 1Hz, 0.5Hz, 0.25Hz, 0.1Hz, respectively. For stroboscopic (50%), bright light time was 6001ux, dark time was 0, and 48 2W old guinea pigs in group.B of the control group were randomly divided into 3 different luminance stroboscopic groups and 1 control groups (n=12). The frequency of stroboscopic group was 0.5Hz (1s dark, 1s bright) frequency with 0-600lux, 0-300lux and 300-600lux brightness alternately stroboscopic, and the control group was 6. 001ux continuous luminance lighting.AB two study groups all experimental guinea pigs were used the same time control switch to control the lighting time as normal resting rhythm 6:00-18:00.C group 36 2W old guinea pigs randomly divided into 2 different rhythm stroboscopic groups and 1 control groups (n=12). The stroboscopic group was flicker with 0.5Hz frequency 0-600lux illumination, stroboscopic exposure and stop stroboscopic The time was 12h/12h (normal resting rhythm 6:00-18:00) and 0.5h/0.5h two kinds of rhythms respectively. The control group was illuminated with 600lux continuous illumination, the light time was from 6:00 to 18:00, and the same as the 12h/12h stroboscopic group. The above 3 experimental groups all used 505nm wavelength light emitting diodes, and the illumination measuring instrument was used to measure instant illumination brightness, each group of guinea pigs of each group was 2 W recorded the diopter, the length of the eye axis and the radius of the curvature of the cornea. The record was 0W for the first day of measurement, and the continuous observation time was drawn by the GRAPHPADPRISM Version5.0 software for 12w., and the SPSS17.0 was statistically analyzed. The double binocular difference was paired t test. The left eye parameters of each guinea pig were further analyzed, and the difference between the groups was adopted. Factor analysis of variance and multiple comparisons showed that P0.05 was statistically significant.
Results: there was no significant difference in biological parameters between the guinea pigs before the experiment (P0.05). There was no significant difference between the two groups (P0.05). As time extended, the frequency of flash groups in the three groups of the three study groups were all offset to myopia and eye axis extension (P0.05) compared with the control group. The radius of curvature of the cornea of each group was all in the whole experimental process. There was no statistical difference between each group (P0.05) at different time points (P0.05).12w. The myopia diopter and eye axis length of each group were compared statistically. The myopia and ocular axis of 0.5Hz stroboscopic group in group A were the most obvious, and the diopter changes in different luminance groups in B group and eye axis lengthening 0-6001ux0-3001ux300-6001ux control In group.C, the change of flexion and axial length of 0.5h/0.5h12h/12h in different rhythm groups were compared.
Conclusions: (1) long exposure to a specific low frequency flash can stimulate the overdevelopment of the eyeball and induce the formation of axial myopia in guinea pigs. There is a difference between the different stimuli frequency and the most significant effect of 0.5Hz frequency flash. (2) the intensity of low frequency flash or the amplitude of light and shade helps to reduce the degree of refractive development. (3) stroboscopic exposure Rhythms may affect the overdevelopment of the eyeball. The phenomenon of abnormal development of the eyeball in guinea pigs suggests that the central nervous regulation mechanism may be involved.
The second part is the effect and mechanism of 0.5Hz flash on guinea pig eyeball structure and function.
Objective: To investigate the effects of sustained 0.5Hz frequency flash exposure on visual transmission function and ultrastructure of retinal sclera in guinea pigs.
Objects and methods: 36 male 2W old male guinea pigs were randomly divided into stroboscopic and control group (n=18). Stroboscopic light flicker was given at the frequency of 0.5Hz (50%, 1s light, 1s dark). The illumination of the cage was measured by the illuminance measuring instrument and the illumination fluctuation was 0-6001ux before the experiment, and the control group adjusted the cage light through the controller knob. 505nm wavelength light emitting diode was used in the continuous 6001ux.2 group. The above 2 groups were controlled by the same control switch. The control light time was normal day and night 12h rotation. After 3 weeks of 6:00-18:00. experiment, 6 guinea pigs of two guinea pigs were selected to separate the retina and sclera tissue, and the Western-blot observation protein was observed. The difference in the expression of the horizontal retina and posterior sclera MMP-2. The other 24 guinea pigs were measured by the electroretinogram (F-ERG) and oscillatory potential (Ops) in the guinea pig before and after the experiment. After the experimental 12W, all guinea pigs were killed and the eyeball was taken out to measure the diameter of the eye water, the vertical diameter and the front and back diameter. The pathological light and the electron microscope view were measured. The ultrastructure of retina and sclera in the posterior polar part of the eyeball was compared with the control group. The GRAPHPADPRISM Version5.0 software was used to chart, and the SPSS17.0 was statistically analyzed. The difference between the groups was determined by t test, and the difference of P0.05 was statistically significant.
Results: compared with the control group, the expression of retina retina and MMP-2 in the retina of each guinea pig increased (P0.05). At 12W, the a wave latency of F-ERG in stroboscopic group was prolonged (P0.05) compared with the control group, and there was no statistical difference between the amplitude of a wave and the latency and amplitude of B wave (P0.05), and there was no statistical significance (P0.05) between the two groups of Ops. The horizontal diameter, vertical diameter and anterior and posterior diameter of the eyeball were 0.89 + 0.3mm, 0.69 + 0.2m and 0.96+0.3mm respectively. The difference was statistically significant (P0.05). The histological observation: the scleral fiber of the light microscope flash group appeared dilatation, the outer segments of the retinal photoreceptor layer were arranged in disorder under electron microscope, and more deformed and exfoliated outer segment membranes were visible. There was no obvious abnormality in the control group.
Conclusion: the expression of visual protein in the 1. frequency flashing induced guinea pig myopia model suggests that the abnormal light perception information of the cone cells may be the increase of the scleral MMP-2 in the starting link of the ocular axis.2. stroboscopic. The relationship between the eye axis of the guinea pig and the eye axis of the guinea pig is worthy of further exploring the overdevelopment of the eyeball induced by.3. continuous frequency flash stimulation. Affect the ultrastructure and conduction function of retinal photoreceptors.
【學(xué)位授予單位】：復(fù)旦大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R778.1

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相關(guān)期刊論文 前1條

1 王紅;莊康;陶遠;王珍紅;李鏡海;;頻閃光對發(fā)育期豚鼠近視的影響[J];環(huán)境與健康雜志;2007年06期

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本文編號：1990382