本文選題：早產(chǎn)兒視網(wǎng)膜病變 + 氧誘導(dǎo)視網(wǎng)膜病變�。� 參考：《第四軍醫(yī)大學(xué)》2013年博士論文

【摘要】：研究背景 早產(chǎn)是一種常見的臨床現(xiàn)象，導(dǎo)致新生兒較高的死亡率和發(fā)病率。臨床證據(jù)表明，早產(chǎn)兒發(fā)生視覺損害和眼部疾病的風(fēng)險明顯高于足月兒。早產(chǎn)可影響視網(wǎng)膜血管系統(tǒng)的結(jié)構(gòu)和功能發(fā)育，從而使得早產(chǎn)兒罹患血管性疾病的閾值降低，臨床上最為常見的表現(xiàn)是早產(chǎn)兒視網(wǎng)膜病變（ROP）。ROP是一種復(fù)雜的視網(wǎng)膜新生血管性疾病，因早產(chǎn)兒的血管發(fā)育異常所致。視網(wǎng)膜血管化越不成熟，發(fā)生嚴(yán)重ROP的風(fēng)險就越大。小胎齡和低出生體重是公認(rèn)的發(fā)生ROP的最重要的危險因素。但是，作為一種多因素致病性疾病，ROP的病因和病理機制目前尚未闡明。 大鼠氧誘導(dǎo)視網(wǎng)膜病變（OIR）模型廣泛用于ROP的實驗研究中，其應(yīng)用基礎(chǔ)為新生大鼠發(fā)育不成熟的視網(wǎng)膜血管系統(tǒng)與人類早產(chǎn)兒極其相似。然而，OIR采用的是足日齡生產(chǎn)的正常新生大鼠，它們出生時和出生后的病理生理狀態(tài)與早產(chǎn)兒有所不同。在新生大鼠中模擬早產(chǎn)兒出生后生長受限的研究發(fā)現(xiàn)，出生后的生長發(fā)育情況決定了幼鼠正常視網(wǎng)膜血管化的速度和OIR中異常血管化的程度。理論上，早產(chǎn)大鼠可在一定程度上模擬人類早產(chǎn)兒出生時的不成熟以及出生后的發(fā)育不良經(jīng)歷，能為研究“早產(chǎn)”這一因素在ROP中的作用提供一個良好的平臺。雖然早產(chǎn)大鼠模型已被廣泛用于研究人類發(fā)育相關(guān)性疾病，但目前有關(guān)早產(chǎn)大鼠的視網(wǎng)膜發(fā)育特點以及早產(chǎn)對大鼠OIR的影響尚無報道。 目的 1.觀察早產(chǎn)大鼠的神經(jīng)視網(wǎng)膜、視網(wǎng)膜淺層毛細(xì)血管的結(jié)構(gòu)以及視網(wǎng)膜功能的發(fā)育特點； 2.觀察早產(chǎn)大鼠在氧誘導(dǎo)條件下的視網(wǎng)膜血管病變情況及視網(wǎng)膜中血管內(nèi)皮生長因子（VEGF）和胰島素樣生長因子1（IGF-1）的表達水平。 方法 1.于Sprague Dawley（SD）大鼠孕19天時行剖宮產(chǎn)，建立早產(chǎn)大鼠模型，觀察空氣中飼養(yǎng)的早產(chǎn)幼鼠的一般情況及體重變化。同時設(shè)正常足日齡生產(chǎn)的大鼠作為對照。 2.分別于幼鼠出生后4、7、10、14天行視網(wǎng)膜切片、HE染色及熒光標(biāo)記的isolectin B4視網(wǎng)膜血管染色、鋪片，觀察幼鼠視網(wǎng)膜各層細(xì)胞和淺層血管形態(tài)，測量神經(jīng)視網(wǎng)膜各層厚度和未血管化區(qū)面積；分別于幼鼠出生后14、21、28、35天行視網(wǎng)膜電圖（ERG）檢測，記錄各波的波幅和潛伏期。 3.將新生大鼠在出生后6h內(nèi)置于80%/21%濃度、24h交替的動物氧箱中，維持2周，于出生后15天返回空氣中飼養(yǎng)，建立大鼠OIR模型；分別于出生后14、18、22天行熒光標(biāo)記的isolectin B4視網(wǎng)膜血管染色、鋪片，觀察視網(wǎng)膜血管形態(tài)，測量視網(wǎng)膜無血管區(qū)及新生血管區(qū)面積；采用反轉(zhuǎn)錄多聚酶鏈?zhǔn)椒磻?yīng)（RT-PCR）檢測視網(wǎng)膜中VEGF、IGF-1mRNA的表達水平；于出生后18天行視網(wǎng)膜切片、HE染色，計數(shù)突破視網(wǎng)膜內(nèi)界膜的血管內(nèi)皮細(xì)胞數(shù)量。 結(jié)果 1.剖宮產(chǎn)手術(shù)的成功率以及早產(chǎn)幼鼠的存活率較高。出生后3周內(nèi)，早產(chǎn)幼鼠的體重低于足日齡幼鼠（P0.001）。 2.早產(chǎn)幼鼠的視網(wǎng)膜各層細(xì)胞和淺層血管形態(tài)及發(fā)育趨勢與對照組相似；出生后早期，早產(chǎn)幼鼠的視網(wǎng)膜厚度大于對照組（P0.05），2周時趨于一致；2周內(nèi)早產(chǎn)幼鼠的視網(wǎng)膜無血管區(qū)明顯大于對照組（P0.001），2周時兩組的視網(wǎng)膜血管化均已完成；出生2周后，早產(chǎn)鼠的ERG波幅和潛伏期與對照組相似。 3.波動氧干預(yù)后，早產(chǎn)幼鼠的視網(wǎng)膜一級血管迂曲程度與對照組相似；早產(chǎn)幼鼠14和18天的視網(wǎng)膜無血管區(qū)及18天的視網(wǎng)膜新生血管區(qū)面積大于對照組（P0.001），早產(chǎn)幼鼠22天時殘留的視網(wǎng)膜血管病變較對照組更多（P0.01）。早產(chǎn)幼鼠14、18、22天的視網(wǎng)膜中VEGF mRNA的表達高于對照組，，IGF-1mRNA的表達無明顯差異。 結(jié)論 剖宮產(chǎn)誘導(dǎo)的早產(chǎn)對幼鼠的體重增長及神經(jīng)視網(wǎng)膜、視網(wǎng)膜血管發(fā)育產(chǎn)生負(fù)性影響。早產(chǎn)幼鼠不成熟的視網(wǎng)膜血管系統(tǒng)更易遭受氧氣的損傷，導(dǎo)致更嚴(yán)重而持久的視網(wǎng)膜病變。這些研究結(jié)果將為進一步在動物模型中研究各種危險因素在ROP發(fā)生過程中的“多重作用”提供基礎(chǔ)數(shù)據(jù)。
[Abstract]:Background of the study

Premature delivery is a common clinical phenomenon , leading to higher mortality and morbidity in neonates . The clinical evidence suggests that preterm infants have a higher risk of visual impairment and eye disease than in term infants . Early birth can affect the structural and functional development of retinal vasculature , which is the most important risk factor for preterm infants . However , as a multi - factor pathogenicity disease , the etiology and pathological mechanism of ROP has not yet been elucidated .

In the experimental study of rat ' s oxygen - induced retinopathy ( OIR ) model widely used in ROP , the application of OIR is very similar to that of preterm infants .

Purpose

1 . To observe the structure of retinal and retinal shallow capillary vessel and the development of retinal function in preterm rats .


2 . The expression level of vascular endothelial growth factor ( VEGF ) and insulin - like growth factor 1 ( IGF - 1 ) in the retina of preterm rats under oxygen - induced conditions were observed .

method

1 . In Sprague Dawley ( SD ) rats , a model of preterm rats was established to observe the general condition and body weight change of preterm young rats fed in air .

2 . At 4 , 7 , 10 and 14 days after birth , retinal slices , HE staining and fluorescent labeled isolectin B4 were stained , and the thickness of retinal layers and the area of non - vascularization area were measured .
Electroretinogram ( ERG ) was detected at 14 , 21 , 28 and 35 days after birth respectively , and the amplitude and latency of each wave were recorded .

3 . The neonatal rats were placed in 80 % / 21 % concentration at 6 h after birth , in an alternate animal oxygen tank for 2 weeks , returned to air for 15 days after birth , and the rat OIR model was established ;
At 14 , 18 and 22 days after birth , the retinal vessels were stained with the fluorescent labeled isolectin B4 , and the retinal vessel morphology was observed and the area of retinal blood vessel and neovascularization was measured .
The expression level of VEGF and IGF - 1 mRNA in the retina was measured by reverse transcription polymerase chain reaction ( RT - PCR ) .
The retinal section and HE staining were performed on 18 days after birth , and the number of vascular endothelial cells was counted .

Results

1 . The success rate of the caesarean operation and the survival rate of the preterm young rats were higher . In the 3 weeks after birth , the weight of the premature infant was lower than that of the young mouse ( P0.001 ) .

2 . The morphology and developmental tendency of retinal layer cells and superficial layers of early mouse were similar to those of the control group .
The retinal thickness of early and early postnatal mice was greater than that of control group ( P0.05 ) , and tended to be consistent in 2 weeks .
There was no significant difference between the two groups ( P 0.001 ) , and the retinal vascularization in two groups was completed in 2 weeks .
After 2 weeks of birth , the ERG amplitude and latency of the preterm mice were similar to those of the control group .

3 . After the intervention of the fluctuating oxygen , the degree of the retinal primary vessel was similar to that of the control group .
Compared with the control group ( P0.01 ) , the expression of VEGF mRNA was higher in the retina without vascular and 18 days earlier than that in the control group ( P0.01 ) . The expression of VEGF mRNA in the retina of early - birth young rats was higher than that in the control group .

Conclusion

The results of these studies will provide basic data for further study of " multiple effects " of various risk factors in the process of ROP .
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2013
【分類號】：R722.6

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