色光對豚鼠鞏膜重塑眼球生長和人眼調(diào)節(jié)系統(tǒng)的作用研究
發(fā)布時間:2018-05-01 03:00
本文選題:色光 + 眼球生長; 參考:《復旦大學》2011年博士論文
【摘要】:近視眼是世界范圍內(nèi)最常見的眼部異常之一,探索近視眼發(fā)病的因素及其機制,以指導有效的防治,是當前研究的熱點和難點。眼球的生長發(fā)育依賴于視覺信號的輸入并對其產(chǎn)生反饋性調(diào)控,以達到使外界視覺物體在視網(wǎng)膜上清晰成像的目的。視覺信號的不同性質(zhì)及眼屈光介質(zhì)客觀存在的不規(guī)則因素等,可導致視網(wǎng)膜產(chǎn)生模糊影像即視網(wǎng)膜離焦。光環(huán)境中不同光譜性質(zhì)形成的視覺信號可能通過作用于眼球局部(視網(wǎng)膜-脈絡膜-鞏膜)以及調(diào)節(jié)系統(tǒng),進而引起屈光狀態(tài)的變化。針對視覺物體或光環(huán)境中特定波長色光的干預,將有望為臨床干預屈光不正提供新的思路。 第一部分 色光對豚鼠鞏膜重塑與眼球生長的作用 第一節(jié)色光對豚鼠屈光發(fā)育及脈絡膜和鞏膜形態(tài)的作用 目的:探討430nm短波長光和530nm長波長光對豚鼠眼球屈光發(fā)育的作用及脈絡膜和鞏膜形態(tài)的變化。 方法:30只2~3周齡健康豚鼠,隨機分為三組,分別飼養(yǎng)于530nm單色光照,430nm單色光照及白色光照環(huán)境(色溫5000K),各光照環(huán)境有效光子量均為3×10-4μmol/cm3光照周期為12/12h。在光照前、光照后4周及8周進行屈光度和眼軸各部分長度測量。光照8周后每組隨機處死5只豚鼠,取眼球行光鏡和透射電鏡檢查。對各組不同時間點的數(shù)據(jù)采用重復測量數(shù)據(jù)的方差分析,以P0.05為有統(tǒng)計學意義。 結(jié)果:1)光照4周后,530nm光照組較白光組形成-0.08±0.41D近視(P=0.725),430nm光照組較白光組形成1.72±0.78D遠視(P0.001);光照8周后,530nm光照組較白光組形成-1.20±0.40D近視(P0.001),430nm光照組較白光組形成2.15±0.87D遠視(P0.001)。2)光照4周后,530nm光照組玻璃體腔長度增加0.15±0.08 mm,較白光組多增長0.05±0.12mm(P=0.326),430nm光照組玻璃體腔長度增加0.02±0.05 mm,較白光組少增加0.09±0.07 mm(P=0.056);光照8周后,530nm光照組玻璃體腔較白光組多增加0.12±0.11 mm(P=0.036);430nm光照組玻璃體腔較白光組少增加0.11±0.08 mm(P=0.006)。各組前節(jié)長度和晶狀體厚度隨光照時間增加,各時間點各組間未見統(tǒng)計學差異(P0.05)。3)光照后8周,530nm光照組鞏膜膠原疏松,排列欠整齊,成纖維細胞處于相對靜止狀態(tài),430nm光照組鞏膜膠原纖維排列整齊緊密,成纖維細胞內(nèi)質(zhì)網(wǎng)擴張,合成代謝旺盛。4)脈絡膜厚度在530nm光照組、430nm組和白光組分別為40.15±10.12μm、41.71±8.2μm和41.29±9.7μm,差異無統(tǒng)計學意義。鞏膜的厚度在530nm光照組顯著小于白光組(93.91±7.1μm vs.97.46±12.3μm,P=0.008),430nm光照組和白光組之間則無顯著性差異(101.32±10.2μm vs.97.46±12.3μm,P=0.057)。 結(jié)論:不同波長的單色光干預可使豚鼠屈光發(fā)育狀態(tài)發(fā)生變化,長波長光可誘導相對性近視,短波長光可誘導相對性遠視。眼球?qū)Σ煌ㄩL色光產(chǎn)生的反應,以鞏膜變化引起的眼球生長速度改變?yōu)橹。長波長光較白光使豚鼠鞏膜變薄,短波長光與白光對鞏膜厚度的影響無顯著差異。不同波長色光對鞏膜性質(zhì)的影響有待進一步研究。 第二節(jié)色光對豚鼠鞏膜生物力學性質(zhì)和細胞外基質(zhì)成分的作用 目的:研究430nm波長光和530nm波長光光照對豚鼠鞏膜生物力學性質(zhì)和細胞外基質(zhì)成分的作用。 方法:光照后4周及8周時各組處死12只豚鼠納入研究,其中6只豚鼠的后極部鞏膜用于鞏膜蠕變量測量?v向剪取2mm×6mm鞏膜條帶,安裝到CMT 6000電子萬能試驗機,經(jīng)過預拉伸試驗后,在0.05N的載荷下,進行20分鐘鞏膜條帶蠕變過程。蠕變量的計算為(L′-L)/L×100%(L′為800s時的形變量,L為200s時的形變量)。每組各6只豚鼠的一眼用于實時熒光定量逆轉(zhuǎn)錄聚合酶鏈反應,檢測鞏膜組織中Ⅰ型膠原蛋白、aggrecan和MMP-2、TIMP-2的mRNA水平表達情況。另一眼用于免疫印跡法蛋白定量,檢測鞏膜組織中Ⅰ型膠原蛋白和aggrecan的蛋白水平的表達情況。同一時間點上三組之間比較采用單因素方差分析(ANOVA), P0.05為差異有統(tǒng)計學意義。 結(jié)果:1)光照后4周和8周,530nm組鞏膜蠕變量均大于白光組(4w,P=0.01;8w,P=0.042);光照后4周時430nm組鞏膜蠕變量小于白光組(P=0.021),8周時430nm組與白光組差異無統(tǒng)計學意義(P=0.067)。2)Real-time PCR檢測發(fā)現(xiàn):與白光組比較,光照后4周及8周,530nm組鞏膜α1(Ⅰ) Collagen mRNA水平降低(4w,P=0.023;8w,P=0.048),430nm組鞏膜α1(Ⅰ) Collagen mRNA水平無顯著差異(4w,P=0.927,8w,P=0.910);530nm組鞏膜組織中aggrecan mRNA水平較白光組降低(4w,P=0.037;8w,P=0.015),430nm組鞏膜中aggrecan mRNA水平在4周時較白光組升高(P=0.047),8周時與白光組無顯著差異(P=0.210);530nm組鞏膜組織中MMP-2 mRNA水平在光照4周時較白光組升高(P=0.007),8周時升高不明顯(P=0.815),TIMP-2 mRNA水平較白光組無顯著變化(P=0.524,P=0.072),430nm組鞏膜中MMP-2 mRNA水平與白光組比較無統(tǒng)計學差異(4w,P=0.947;8w,P=0.510),TIMP-2 mRNA水平與白光組無顯著差異(P=0.375,P=0.810)。3)Western-blot檢測Ⅰ型膠原和aggrecan蛋白表達結(jié)果與RT-PCR結(jié)果一致。 結(jié)論:不同波長色光引起豚鼠鞏膜蠕變性質(zhì)改變,530nm波長光較430nm及白光使鞏膜更易于延展。不同波長光致鞏膜細胞外基質(zhì)成分改變,530nm波長光照下鞏膜Ⅰ型膠原蛋白、aggrecan表達水平降低,參與分解代謝的酶MMP-2表達水平上調(diào)。色光調(diào)控豚鼠眼球生長與鞏膜細胞外基質(zhì)重塑有關,機制有待進一步研究。 第二部分 色光對近距調(diào)節(jié)反應及調(diào)節(jié)系統(tǒng)模糊敏感度的影響 目的:研究不同波長色光對近距靜態(tài)調(diào)節(jié)反應及調(diào)節(jié)系統(tǒng)模糊敏感度的影響。 方法:入選12名被試,平均年齡為25.5±3.4歲(20-30歲)。所有入選被試單眼屈光不正的等效球鏡度數(shù)低于-6.00 D,最佳矯正視力達20/20或以上,無雙眼視功能異常,色覺正常。在實驗過程中均用軟性角膜接觸鏡矯正屈光不正,戴鏡后等效球鏡度數(shù)在±0.25 D之內(nèi)。本實驗僅采集右眼數(shù)據(jù)。被試右眼等效球鏡值范圍為0D至-4D,平均為-1.23±1.49 D。采用3×3陣列空間頻率為12 c/deg的Snellen E字母作為注視視標,整個視標呈現(xiàn)的視場為2°。根據(jù)1931CIE-XYZ標準色度系統(tǒng)的參考值,設置三種色度的視標,字體與背景顏色分別為黑-白(B-W),藍-黃(B-Y)和紅-綠(R-G)。藍色和紅色的主波長分別為440nm和650nm,背景色分別為兩者的拮抗色,主波長分別為580nm和510nm。三種視標的平均亮度相同。用Badal系統(tǒng)呈現(xiàn)3D調(diào)節(jié)刺激水平。用近紅外開放視野自動驗光儀(Grand Seiko WAM 5500)測量調(diào)節(jié)反應。每隔0.2s連續(xù)采集調(diào)節(jié)微波動數(shù)據(jù),連續(xù)記錄調(diào)節(jié)反應20秒,以均方根值(r.m.s.)作為調(diào)節(jié)微波動幅度。在3D調(diào)節(jié)刺激水平上以0.1 D的梯度改變調(diào)節(jié)刺激水平,監(jiān)測調(diào)節(jié)反應,采用t檢驗法判定調(diào)節(jié)反應變化點,將能引起調(diào)節(jié)反應相對于基礎水平發(fā)生變化的調(diào)節(jié)刺激最小改變量作為模糊感知閾值。采用重復測量方差分析統(tǒng)計檢驗注視三種視標時的調(diào)節(jié)反應、調(diào)節(jié)反應均方根.值及模糊感知閾值的差異。P0.05具有統(tǒng)計學意義。 結(jié)果:1)在3D調(diào)節(jié)刺激水平上黑-白、藍-黃及紅-綠視標誘發(fā)的平均靜態(tài)調(diào)節(jié)反應分別為2.37±0.18D,2.06±0.20D,2.16±0.21D,黑-白視標誘導的靜態(tài)調(diào)節(jié)反應高于紅-綠視標(P=0.044)和藍-黃視標(P=0.010);紅-綠視標下的調(diào)節(jié)反應高于藍-黃視標,差值為0.11±0.03 D(p=0.008)。2)在3D調(diào)節(jié)刺激水平,注視黑-白,藍-黃,紅-綠視標的調(diào)節(jié)反應均方根平均值分別為0.173±0.027D,0.164±0.035D,0.206±0.014D,黑-白與藍-黃視標無顯著差異(P=0.171),紅-綠視標顯著大于黑-白(P=0.025)與藍-黃視標(P=0.021)。3)在3D調(diào)節(jié)刺激水平,注視黑-白、紅-綠及藍-黃視標的平均模糊感知閾值分別為0.135±0.023D,0.133±0.041D,0.203±0.050D。黑-白視標與藍-黃視標無顯著性差異(P=0.836),紅-綠視標顯著大于黑-白(P=0.030)和藍-黃視標(P=0.028)。結(jié)論:調(diào)節(jié)系統(tǒng)對紅-綠顏色視標的模糊敏感度和穩(wěn)定性較藍-黃和黑-白視標低。不同波長色光可影響近距靜態(tài)調(diào)節(jié)反應水平及調(diào)節(jié)系統(tǒng)模糊敏感度和穩(wěn)定性。不同波長色光對調(diào)節(jié)系統(tǒng)模糊敏感度和穩(wěn)定性的影響趨勢一致。短波長視錐細胞和相應色覺信號的參與可能與提高調(diào)節(jié)系統(tǒng)模糊敏感度及穩(wěn)定性有關,有待進一步研究。
[Abstract]:The development of visual signal depends on the input of visual signal and its mechanism to guide the effective prevention and control . It is the focus and difficulty of the current research . The development of eyeball depends on the input of visual signal and its feedback control , so as to achieve the purpose of making the external visual object clear image on the retina . The visual signal of different spectral properties in the light environment can lead to the change of refractive state . For the intervention of specific wavelength light in the visual object or light environment , it is hopeful to provide a new idea for clinical intervention .
the first portion
Effects of color light on scleral remodeling and eyeball growth in guinea pigs
The effect of the first color light on the refractive development of guinea pigs and the shape of choroidotes and sclera
Objective : To investigate the effect of 430 nm short wavelength light and 530 nm long wavelength light on the optic development of guinea pig ' s eyeball and the changes of choroidal and scleral morphology .
Methods : 30 healthy guinea pigs from 2 to 3 weeks old were randomly divided into three groups : 530 nm monochromatic light , 430 nm monochromatic light and white light environment ( color temperature 5000K ) . The effective photon quantity of each illumination environment was 3 脳 10 - 4 渭mol / cm ~ 3 illumination period of 12 / 12 h . After 8 weeks of illumination , 5 guinea pigs were sacrificed at random .
Results : 1 ) After 4 weeks of illumination , the 530 nm light group formed - 0.08 鹵 0.41D myopia ( P = 0.725 ) compared with the white light group , and the light group of 430 nm had a far vision ( P < 0.001 ) than that of the white light group .
After 4 weeks of illumination , the vitreous cavity length of the 530 nm light group increased by 0.15 鹵 0.08 mm , the length of vitreous cavity increased by 0.05 鹵 0.12 mm ( P = 0.326 ) , the length of vitreous cavity increased by 0.02 鹵 0.05 mm and 0.09 鹵 0.07 mm ( P = 0.056 ) .
After 8 weeks of illumination , the vitreous cavity of the 530 nm light group increased by 0.12 鹵 0.11 mm ( P = 0.036 ) .
The thickness of sclera was significantly lower than that in white light group ( 93.91 鹵 7.1 渭m vs . 97.46 鹵 12.3 渭m , P = 0.008 ) .
Conclusion : Single - color light intervention at different wavelengths can change the refractive state of guinea pig , and the long - wavelength light can induce relative myopia . The short - wavelength light can induce relative hyperopia . The effects of different wavelengths of light on scleral thickness are not significant . The effects of different wavelengths of light on scleral properties are to be further studied .
The effect of the second color light on the biomechanical properties and extracellular matrix composition of the sclera of guinea pig
Objective : To study the effects of 430 nm wavelength light and 530 nm wavelength light on the biomechanical properties and extracellular matrix composition of guinea pig sclera .
Methods : Twelve guinea pigs were sacrificed at 4 and 8 weeks after illumination . Six guinea pigs were taken for scleral creep measurement . After pre - stretching test , the creep process of scleral strip was carried out under 0.05 N loading . The creep quantity was calculated as ( L ' - L ) / L 脳 100 % ( shape variable at L ' = 800s , shape variable at L = 200s ) . The expression of type I collagen , aggrecan and MMP - 2 and TIMP - 2 mRNA in scleral tissues was detected by a single - factor quantitative reverse transcription polymerase chain reaction ( RT - PCR ) .
Results : 1 ) The scleral creep of 530 nm group was greater than that of white light group ( 4w , P = 0.01 ) at 4 and 8 weeks after illumination .
8w,P=0.042)錛,
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