本文選題：半胱氨酸天冬氨酸蛋白酶3 + 形覺剝奪性弱視　； 參考：《南華大學(xué)》2013年碩士論文

【摘要】：目的 研究細(xì)胞凋亡因子半胱氨酸天冬氨酸蛋白酶3（Cysteine protease protein32KD，Caspase-3）在形覺剝奪性弱視大鼠視皮層及外側(cè)膝狀體的表達(dá)，探討其與弱視發(fā)病機(jī)理的關(guān)系，期望能對弱視的治療產(chǎn)生積極的影響。 方法 1.剛出生的健康SD（Sprague Dawley）大鼠40只，飼養(yǎng)14天后隨機(jī)選擇20只行單眼眼瞼縫合術(shù)為單眼形覺剝奪組（Monocular Deprivation，MD），正常組（Normal Postnatal， NP）不做特殊處理。根據(jù)飼養(yǎng)天數(shù)分別在14天、21天、45天、120天處死動物。MD組即分為MD14、MD21、MD45、MD120；NP組分為NP14、NP21、NP45、NP120，每組各5只。 2.將NP21、MD21組的大鼠稱重麻醉后行P-VEP（Pattern Visual EvokedPotential）檢測，記錄P100波的潛伏期和波幅值。 3.采用HE（Hematoxylin Eosin）染色和免疫組化法以及圖像分析系統(tǒng)對正常大鼠和單眼形覺剝奪弱視大鼠視皮層、外側(cè)膝狀體中Caspase-3免疫陽性神經(jīng)元進(jìn)行定位觀察，，并根據(jù)測定的平均灰度值定量研究其變化。 結(jié)果 1.P-VEP檢測：MD21組大鼠弱視眼的P-VEP P100波峰潛伏期檢測值較NP21組同側(cè)眼顯著延長(P0.01)，波幅明顯降低(P0.01)，提示MD21組弱視眼的視覺電生理已改變，表明可以通過單眼瞼縫合術(shù)成功構(gòu)建形覺剝奪弱視模型。 2.（1）HE染色：NP45大鼠大腦視皮層神經(jīng)元的結(jié)構(gòu)正常，細(xì)胞輪廓清晰，可清晰地顯示神經(jīng)元的形態(tài)結(jié)構(gòu)及染色，MD45剝奪組神經(jīng)元細(xì)胞排列不整齊，胞膜結(jié)構(gòu)不清，神經(jīng)元變性，部分神經(jīng)元細(xì)胞核固縮，呈強(qiáng)嗜堿性，胞漿深染，結(jié)構(gòu)不清。 （2）Caspase-3免疫陽性神經(jīng)元在視皮層中的表達(dá)：Caspase-3經(jīng)染色后呈棕黃色，陰性細(xì)胞為藍(lán)色。出生后14天（P14）Caspase-3的平均灰度值在NP14和MD14兩組無顯著性差異（P=0.074），而出生后21天、45天、120天（P21、P45、P120），Caspase-3免疫反應(yīng)產(chǎn)物在MD組大鼠視皮層中表達(dá)密集，NP組表達(dá)產(chǎn)物較前基本無改變。P21、P45、P120三個時段MD組平均灰度值均較同時期NP組顯著性升高（P＜0.05）。在MD組MD21和MD45組間無顯著性差異（P0.05），MD21和MD45與MD14、MD120之間均有差異顯著性（P＜0.05）。 （3）Caspase-3免疫陽性神經(jīng)元在外側(cè)膝狀體的表達(dá)：在MD組大鼠LGN中顆粒狀物分布密集，但不同時期其密度也不同。NP組大鼠LGN的Caspase-3表達(dá)產(chǎn)物分布稀疏，P14：MD組和NP組兩組無顯著性差異(P=0.168)，P21、P45、P120三個時段MD組均較同時期NP組平均灰度值顯著性升高(P＜0.05)。在MD組MD14、MD21、MD45和MD120組間兩兩比較均有差異顯著性（P＜0.05）。 （4）MD組Caspase-3在大鼠視皮層及外側(cè)膝狀體的表達(dá)量隨剝奪時間變化，其在視覺發(fā)育關(guān)鍵期形覺剝奪開始時增加，到早中期持續(xù)高表達(dá)，視覺發(fā)育關(guān)鍵期結(jié)束后稍降低，但仍高于形覺剝奪開始時。 結(jié)論 1.在大鼠視覺發(fā)育關(guān)鍵期的早期，應(yīng)用單側(cè)眼瞼縫合術(shù)即使剝奪是短暫的（7天）可以成功建立形覺剝奪性弱視模型。 2.大鼠單眼形覺剝奪會引起視皮層神經(jīng)細(xì)胞發(fā)生形態(tài)學(xué)改變，引起細(xì)胞類凋亡。 3.大鼠單眼形覺剝奪后，Caspase-3在視皮層和外側(cè)膝狀體中的表達(dá)：14天開始增加，21天明顯增高，45天達(dá)到峰值，120天時較前稍下降，但仍高于14天開始剝奪時。 4.單眼形覺剝奪通過引起Caspase-3的表達(dá)增加，進(jìn)而導(dǎo)致神經(jīng)元細(xì)胞凋亡損傷，可能是弱視的發(fā)病機(jī)理之一。
[Abstract]:objective
The expression of cysteine aspartic proteinase 3 (Cysteine protease protein32KD, Caspase-3) in the visual cortex and lateral geniculate body of rats with form deprivation amblyopia is studied to explore the relationship with the pathogenesis of amblyopia, hoping to have a positive effect on the treatment of amblyopia.
Method
1. newly born healthy SD (Sprague Dawley) rats, and 14 days after feeding, 20 single eye eyelid sutures were randomly selected as monocular form deprivation group (Monocular Deprivation, MD). The normal group (Normal Postnatal, NP) did not do special treatment. The animals were killed in 14 days, 21 days, 45 days and 120 days respectively. The.MD groups were divided into MD14, MD21, and 120 days. MD120; NP were NP14, NP21, NP45 and NP120, 5 in each group.
2. the rats in group NP21 and group MD21 were anesthetized and P-VEP (Pattern Visual EvokedPotential) was detected. The latency and amplitude of P100 wave were recorded.
3. the HE (Hematoxylin Eosin) staining, immunohistochemical method and image analysis system were used to locate the Caspase-3 immunoreactive neurons in the visual cortex and the lateral geniculate body of normal rats and monocular deprivation amblyopia rats, and the changes were quantified according to the average gray value measured.
Result
1.P-VEP detection: the detection value of P-VEP P100 wave peak latency in the amblyopia of group MD21 rats was significantly longer than that in the NP21 group (P0.01), and the amplitude of the wave decreased significantly (P0.01), suggesting that the visual electrophysiology of the amblyopia was changed in MD21 group, indicating that the form deprivation amblyopia model could be constructed successfully by monocular eyelid suture.
2. (1) HE staining: the structure of the neurons in the visual cortex of the brain of NP45 rats is normal and the cell outline is clear. The morphological structure and dyeing of the neurons can be clearly displayed. The neuron cells in the MD45 deprivation group are in irregular arrangement, the membrane structure is not clear, the neurons are denatured, and the nuclei of some neurons are strongly basophilic, the cytoplasm is deeply dyed and the structure is not clear.
(2) the expression of Caspase-3 immunoreactive neurons in the visual cortex: Caspase-3 was brown in brown, and the negative cells were blue. The average gray value of 14 days after birth (P14) Caspase-3 had no significant difference in NP14 and MD14 two groups (P=0.074), while 21 days after birth, 45 days, 120 days (P21, P45, P120), Caspase-3 immunoreaction products in MD group rats. The expression in the cortex was dense, and the expression products of NP group had no changes in.P21, P45, and P120 were significantly higher in MD group than that in the same period NP group (P < 0.05). There was no significant difference between the MD21 and MD45 groups in MD group (P0.05), and there was a significant difference between MD21 and MD.
(3) expression of Caspase-3 immunoreactive neurons in the lateral geniculate body: the distribution of granules in LGN of group MD rats was dense, but the density of the Caspase-3 expression products of LGN in group.NP was sparse in different period, and there was no significant difference between group P14:MD and NP group (P=0.168), P21, P45, and P120 groups were all flat compared with the same period. The mean gray value increased significantly (P < 0.05). There was a significant difference between 22 groups in group MD, MD14, MD21, MD45 and MD120 (P < 0.05).
(4) the expression of Caspase-3 in the visual cortex and lateral geniculate body of the MD group changes with the deprivation time. It increases at the beginning of visual development at the critical stage of form deprivation, and continues to express high in the early and middle stages. The critical stage of visual development decreases slightly after the end of visual development, but still higher than the beginning of form deprivation.
conclusion
1. in the early stage of the critical period of visual development in rats, unilateral eyelid suture was applied, even if deprivation was transient (7 days), and the form deprivation amblyopia model could be successfully established.
2. rats with monocular deprivation can cause morphological changes of neurons in the visual cortex and induce cell apoptosis.
The expression of Caspase-3 in the visual cortex and lateral geniculate body after the monocular deprivation of the 3. rats: the increase in the visual cortex and the lateral geniculate body: the increase in the 14 day, the 21 day increase, the peak of the 45 day, and a slight decrease at 120 days, but still higher than the 14 days of deprivation.
4. monocular deprivation can induce neuronal apoptosis by increasing the expression of Caspase-3, which may be one of the pathogenesis of amblyopia.

【學(xué)位授予單位】：南華大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R777.44

【共引文獻(xiàn)】

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