本文選題：糖尿病性白內(nèi)障 + 糖基化終末產(chǎn)物�。� 參考：《瀘州醫(yī)學(xué)院》2014年碩士論文

【摘要】：目的：建立糖尿病大鼠模型，動態(tài)觀察晶狀體的混濁程度、氧化損傷程度、晶狀體內(nèi)糖基化終末產(chǎn)物（AGEs）濃度、糖基化終末產(chǎn)物受體（RAGE）及核因子-κBp65（NF-κBp65）mRNA和蛋白水平的變化，探討AGEs-RAGE軸體系在糖尿病性白內(nèi)障發(fā)生發(fā)展中的作用及機(jī)制。方法：隨機(jī)將無眼部疾病的SD大鼠分成正常組（32只）和糖尿病組（36只），糖尿病組以單次腹腔注射1%鏈脲佐菌素（STZ）溶液65mg/kg誘導(dǎo)糖尿病，正常組注射同等體積的0.1mmol/L檸檬酸鹽緩沖液。3天后空腹（禁食12小時）尾靜脈血糖高于16.6mmol/L為糖尿病成模。每天觀察大鼠一般情況，每兩周檢測記錄血糖、體質(zhì)量，每周麻醉及復(fù)方托吡卡胺滴眼液散瞳后在裂隙燈下觀察晶狀體混濁情況。在實驗4、8、12周時分批處死大鼠，即刻取出雙眼晶狀體。每組各取3枚晶狀體制成石蠟切片，HE染色觀察晶狀體細(xì)胞學(xué)及免疫組化檢測晶狀體上皮細(xì)胞中RAGE、NF-κBp65表達(dá)；5枚晶狀體提取RNA后RT-PCR檢測RAGE mRNA和NF-κBp65mRNA表達(dá)；12枚晶狀體勻漿后檢測AGEs濃度、T-SOD、GSH-PX和CAT活力及MDA含量。結(jié)果：1.注射STZ后大鼠出現(xiàn)糖尿病典型癥狀，血糖明顯高于正常組（t4周=22.84，P=0.000；t8周=26.01，P=0.000；t12周=30.94，P=0.000）。早期糖尿病大鼠多食但體質(zhì)量增加不明顯，隨著病程進(jìn)展逐漸消瘦，這與正常組的體質(zhì)量逐漸增加有顯著差別（t4周=6.54，P=0.000；t8周=20.39，P=0.000；t12周=46.91，P=0.000）。2.正常組大鼠晶狀體始終透明；糖尿病組在實驗4周時部分晶狀體周邊部及前囊膜下出現(xiàn)細(xì)小空泡，隨后空泡向中心擴(kuò)展，瞳孔區(qū)可見團(tuán)塊樣混濁，8周時有皮質(zhì)混濁，此后混濁逐漸累及核周皮質(zhì)及核，至12周時出現(xiàn)晶狀體完全混濁。3. HE染色顯示糖尿病大鼠晶狀體上皮細(xì)胞（LECs）形態(tài)多樣，，胞質(zhì)淡染伴有空泡、胞核縮小；晶狀體纖維排列紊亂，其間有較多囊泡及水裂。4.正常組LECs中幾乎無RAGE表達(dá)、NF-κBp65在胞質(zhì)低表達(dá)，糖尿病組RAGE和NF-κBp65均呈強(qiáng)陽性表達(dá)（RAGE：t4周=90.73，P=0.000；t8周=176.77，P=0.000；t12周=148.67，P=0.000；NF-κBp65：t4周=83.98，P=0.000；t8周=130.61，P=0.000；t12周=195.69，P=0.000），且隨著病程發(fā)展表達(dá)進(jìn)行性增加。糖尿病組晶狀體中RAGEmRNA和NF-κBp65mRNA的表達(dá)均明顯高于正常組（RAGE：t4周=20.36，P=0.000；t8周=29.90，P=0.000；t12周=57.19，P=0.000；NF-κBp65：t4周=11.34，P=0.000；t8周=32.21，P=0.000；t12周=41.03，P=0.000）。5.在實驗4、8、12周時糖尿病組晶狀體中AGEs濃度均高于正常組（t=9.01，P=0.000；t=6.45，P=0.000；t=9.38，P=0.000），且隨著病程延長出現(xiàn)AGEs大量蓄積。6.糖尿病組晶狀體中SOD、GSH-PX、CAT的活力均明顯降低正常組，MDA含量顯著升高（P均小于0.05）。結(jié)論：1.糖尿病大鼠晶狀體內(nèi)AGEs蓄積并與RAGE結(jié)合可增強(qiáng)NF-κB活性；活化的NF-κBp65可上調(diào)RAGE的轉(zhuǎn)錄與合成，并與更多的AGEs作用。2.隨著病程延長，糖尿病大鼠晶狀體內(nèi)生成AGEs增多，抗氧化酶活性降低，晶狀體上皮細(xì)胞氧化損傷加重，晶狀體混濁加重。3.晶狀體內(nèi)AGEs-RAGE軸體系介導(dǎo)的氧化應(yīng)激、非酶糖基化作用中多種信號轉(zhuǎn)導(dǎo)通路并形成級聯(lián)放大反應(yīng)可能是糖尿病性白內(nèi)障發(fā)生發(fā)展的機(jī)制之一，NF-κB是此過程中關(guān)鍵的中間物。
[Abstract]:Objective : To establish a model of diabetic rats , to dynamically observe the degree of opacity , oxidative damage , end - of - end product ( AGE ) , end - product receptor ( RAGE ) and nuclear factor - 魏B ( NF - 魏B ) mRNA and protein levels in diabetic rats .
The expression of RAGE mRNA and NF - 魏B p65mRNA was detected by RT - PCR after extraction of RNA from five lenses .
Results : 1 . The diabetic symptoms of diabetic rats were higher than those in the normal group ( t4 weeks = 22.84 , P = 0.000 ) .
8 weeks = 26.01 , P = 0.000 ;
12 weeks = 30.94 , P = 0.000 ) . In the early diabetic rats , the body mass of the rats was not significantly increased , and the body mass of the normal group increased gradually with the progressive loss of the disease course ( t4 = 6.54 , P = 0.000 ) .
8 weeks = 20.39 , P = 0.000 ;
12 weeks = 46.91 , P = 0.000 ) .
In the diabetic group , small vacuoles appeared in the peripheral part of the lens and the anterior capsule at 4 weeks of the experiment , followed by the extension of the cavity to the center . The area of the pupil was turbid , and the cortex was turbid at 8 weeks . After that , the opacity gradually increased to the cortex and the nucleus , and the lens was completely cloudy at 12 weeks . HE staining showed that the lens epithelial cells ( LECs ) of diabetic rats were varied in shape , and the cytoplasm was light - stained with vacuoles and the nuclei were reduced ;
4 . There was almost no RAGE expression in LECs in the normal group , the low expression of NF - NF - 魏B in the normal group , the low expression of NF - 魏B , the expression of RAGE and NF - 魏B in the diabetic group ( RAGE : t4 = 90.73 , P = 0.000 ) .
8 weeks = 176.77 , P = 0.000 ;
12 weeks = 148.67 , P = 0.000 ;
NF - 魏B P65 : t4 week = 83.98 , P = 0.000 ;
8 weeks = 130.61 , P = 0.000 ;
The expression of RAGEmRNA and NF - 魏B p65mRNA in diabetic group was significantly higher than that in the normal group ( RAGE : t4 = 20.36 , P = 0.000 ) .
8 weeks = 29.90 , P = 0.000 ;
12 weeks = 57.19 , P = 0.000 ;
NF - 魏B P65 : t4 week = 11.34 , P = 0.000 ;
8 weeks = 32.21 , P = 0.000 ;
At the 4th , 8th and 12th week of the experiment , the concentration in the lens of the diabetic group was higher than that in the normal group ( t = 9.01 , P = 0.000 ) .
t=6.45,P=0.000錛

本文編號：1990503