本文選題：胸腺素β4 + 后發(fā)性白內(nèi)障　； 參考：《華中科技大學》2013年碩士論文

【摘要】：目的：后囊混濁（Posteriorcapsuleopaciftcation，PCO）主要由術(shù)后殘留的晶狀體上皮細胞（lensepithelialcells，LECs）的增殖、遷移及上皮間質(zhì)轉(zhuǎn)分化（epithelial mesenchymaltransition，EMT）引起。胸腺素β4（thymosinβ4，Tβ4）是一種重要的肌動蛋白耦聯(lián)蛋白，參與介導多種生物學反應。本實驗通過研究Tβ4對體外培養(yǎng)的人晶體上皮細胞增殖、遷移及上皮間質(zhì)轉(zhuǎn)分化的影響，探討Tβ4在PCO防治中的可能作用。 方法 1.用MTT法檢測Tβ4對LECs增殖的影響。用含不同濃度Tβ4（0ng/ml、1ng/ml、10ng/ml、100ng/ml、1000ng/ml）無血清培養(yǎng)基處理LECs，分別于24h、48h、72h行MTT檢測，測量OD值。 2.細胞劃痕實驗及Transwell小室遷移模型觀察Tβ4對LECs遷移的影響。 （1）劃痕后細胞培養(yǎng)基中加入不同濃度Tβ4（0ng/ml、1ng/ml、10ng/ml、100ng/ml、1000ng/ml）的0.1%牛血清白蛋白DMEM培養(yǎng)基，分別于劃痕后0h、24h、48h選取10個隨機視野拍照記錄劃痕間距（前后拍照位置須一致），比較不同時間點不同濃度的平均遷移距離。 （2）種細胞于上室，下室為不同濃度Tβ4（0ng/ml、1ng/ml、10ng/ml、100ng/ml、1000ng/ml）的20%胎牛血清DMEM培養(yǎng)基，作用24h后比較不同濃度穿過小室的平均細胞數(shù)目。 3.實時定量PCR技術(shù)比較處理72h后Tβ4處理組和正常對照組中E-鈣粘蛋白及α-SMAmRNA表達水平的變化以檢測Tβ4對LECs的轉(zhuǎn)分化作用的影響。 結(jié)果 1.與對照組相比，當濃度為100ng/ml、1000ng/ml時Tβ4能夠明顯促進LECs的增殖，差異有顯著性(P0．05)。相同藥物濃度組隨著作用時間的延長促進作用加強，各時間點相互比較差異亦有顯著性(P0．05)。 2.24h各濃度組（0~1000ng/ml）平均遷移距離分別為：329.6621±47.43715μm,354.2761±52.5730μm,453.5897±46.3792μm,499.6127±46.0270μm,555.4763±71.6129μm;48h各濃度組（0~1000ng/ml）平均遷移距離為：614.6647±97.2274μm,636.1453±649.7268μm,720.5249±41.9510μm,739.9166±85.3627μm,770.9152±62.3144μm。10ng/ml、100ng/ml、1000ng/ml處理組相對于對照組而言，差異有統(tǒng)計學意義（P＜0.05），且細胞遷移距離隨時間和濃度遞增。 Tβ4處理24h后，穿過小室的細胞數(shù)各組（0~1000ng/ml）分別為82±3個,109±6個,128±10個，163±5個,204±11個。10ng/ml、100ng/ml、1000ng/ml處理組相對于對照組而言，差異有統(tǒng)計學意義（P＜0.05），且細胞遷移能力隨濃度遞增。 3.Tβ_4加藥處理72h后LECs表達α-SMA、E-鈣粘蛋白mRNA水平的變化：加藥處理后，α-SMA表達量升高（151.7±5.0）%，E-鈣粘蛋白表達量下降(68.0±8.1)%，差異有統(tǒng)計學意義（P＜0.05）。 結(jié)論：胸腺素β4可促進人晶狀體上皮細胞增殖、遷移及上皮間質(zhì)轉(zhuǎn)分化，其在PCO防治中的可能作用值得作進一步研究。
[Abstract]:Aim: the posterior capsular opacification (PCOs) is mainly caused by the proliferation, migration and epithelial mesenchymal transition of lens epithelial cells (LECs).Thymosin 尾 4(thymosin 尾 4 (T 尾 4) is an important actin coupling protein involved in many biological responses.In this study, we studied the effects of T 尾 4 on the proliferation, migration and epithelial stromal transdifferentiation of cultured human lens epithelial cells in vitro, and explored the possible role of T 尾 4 in the prevention and treatment of PCO.Method1.The effect of T 尾 4 on the proliferation of LECs was detected by MTT assay.LECs were treated with 10ng / ml 10ng / ml 10ng / ml 10ng / ml 10ng / ml 10ng / ml 10ng / ml 10 ng / ml 10 ng / ml 10 ng / ml T 尾 4 / ml 0 ng / ml of different concentrations of T 尾 4mg / ml respectively. The MTT was measured at 24 h / 48h / 72 h, respectively, and OD value was measured.2.The effect of T 尾 4 on LECs migration was observed by cell scratch test and Transwell compartment migration model.After scratch, 0.1% bovine serum albumin (BSA) DMEM medium was added to the cell culture medium with different concentrations of T 尾 4 0 ng / ml 1 ng / ml 10 ng / ml 10 ng / ml 10 ng / ml 100 ng / ml 100 ng / ml 1 000 ng / ml.Ten random visual fields were selected to record the scratch spacing at 0 h 24 h and 48 h after scratch (the position before and after taking pictures should be consistent to compare the average migration distance of different concentrations at different time points.(2) A 20% fetal bovine serum DMEM medium with different concentrations of T 尾 4 0 ng / ml 1 ng / ml 1 ng / ml 10 ng / ml 10 ng / ml 10 ng / ml 100 ng / ml 100 ng / ml 1 000 ng / ml) was used to compare the average number of cells passing through the cell at different concentrations after 24 hours of exposure.3.The changes of Ecadherin and 偽 -SMA mRNA expression in T 尾 4 treated group and normal control group were compared by real time quantitative PCR technique in order to detect the effect of T 尾 4 on the transdifferentiation of LECs.Result1.Compared with the control group, T 尾 4 could significantly promote the proliferation of LECs when the concentration was 100ng / ml or 1000ng / ml, and the difference was significant (P 0.05).The effect of the same drug concentration group was enhanced with the prolongation of the action time, and there were significant differences among the different time points (P 0.05).2.24h鍚勬祿搴︾粍(0~1000ng/ml)騫沖潎榪佺Щ璺濈鍒嗗埆涓，

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