【摘要】：目的:內(nèi)質(zhì)網(wǎng)應(yīng)激在糖尿病視網(wǎng)膜病的發(fā)生和發(fā)展中起到重要作用,而羥甲基戊二酰輔酶A還原酶降解蛋白1(Hrd1)能通過(guò)內(nèi)質(zhì)網(wǎng)相關(guān)蛋白降解途徑穩(wěn)定內(nèi)質(zhì)網(wǎng)內(nèi)環(huán)境從而減少內(nèi)質(zhì)網(wǎng)應(yīng)激的發(fā)生。本實(shí)驗(yàn)通過(guò)基因芯片分析內(nèi)質(zhì)網(wǎng)相關(guān)因子的變化,并觀(guān)察Hrd1在糖尿病鼠視網(wǎng)膜和體外高糖培養(yǎng)人視網(wǎng)膜微血管內(nèi)皮細(xì)胞的表達(dá)變化,分析它在糖尿病視網(wǎng)膜病變發(fā)生和發(fā)展中的作用。 方法腹腔注射鏈脲佐菌素(STZ)(65 mg/kg)制作糖尿病SD大鼠模型,對(duì)照組大鼠腹腔注射同等劑量檸檬酸鈉溶液。于注射后72小時(shí)用鼠尾采血法測(cè)量血糖,7天后以血糖值≥16.7mmol/L認(rèn)定為Ⅰ型糖尿病鼠建模成立,以后每周監(jiān)測(cè)血糖一次。模型成立三個(gè)月時(shí)取大鼠視網(wǎng)膜組織。用自行設(shè)計(jì)實(shí)時(shí)定量PCR芯片分析89個(gè)內(nèi)質(zhì)網(wǎng)相關(guān)因子的表達(dá)變化。用Western blot、免疫熒光檢測(cè)Hrd1在三月糖尿病模型鼠和對(duì)照組大鼠視網(wǎng)膜中的表達(dá)。對(duì)體外培養(yǎng)人視網(wǎng)膜微血管內(nèi)皮細(xì)胞,用高糖培養(yǎng)基干預(yù)48小時(shí)后,用免疫熒光檢測(cè)Hrd1在細(xì)胞的表達(dá)變化。 結(jié)果在視網(wǎng)膜組織中:PCR芯片結(jié)果顯示,在這89個(gè)基因中,三個(gè)月糖尿病模型組有12個(gè)基因mRNA水平發(fā)生了變化,其中Hrd1表達(dá)下降,與正常對(duì)照組相比,差異有統(tǒng)計(jì)學(xué)意義(P0.05);Western blot顯示,三個(gè)月糖尿病模型組Hrd1蛋白質(zhì)表達(dá)水平與對(duì)照組相比下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05);免疫熒光顯示, FITC標(biāo)記的Hrd1在糖尿病鼠視網(wǎng)膜中綠色熒光減弱,表明其蛋白質(zhì)表達(dá)下降,與對(duì)照組相比差異有統(tǒng)計(jì)學(xué)意義(P0.05)。在體外培養(yǎng)血管內(nèi)皮細(xì)胞:免疫熒光檢測(cè)結(jié)果顯示,在高糖培養(yǎng)基干預(yù)后的細(xì)胞中綠色熒光減弱,Hrd1蛋白表達(dá)與對(duì)照組相比顯著下降,差異有統(tǒng)計(jì)學(xué)意義(P0.05)。 結(jié)論:發(fā)生變化的12個(gè)內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)因子參與了糖尿病視網(wǎng)膜病變的發(fā)生和發(fā)展,Hrd1在糖尿病視網(wǎng)膜病變的內(nèi)質(zhì)網(wǎng)應(yīng)激中起到重要作用。
[Abstract]:Objective: endoplasmic reticulum stress plays an important role in the occurrence and development of diabetic retinopathy. However, hydroxymethyl glutaryl coA reductase degrading protein 1 (Hrd1) can stabilize the environment of endoplasmic reticulum through endoplasmic reticulum associated protein degradation pathway, thus reducing the occurrence of endoplasmic reticulum stress. The changes of endoplasmic reticulum (ER) related factors and the expression of Hrd1 in diabetic rat retina and human retinal microvascular endothelial cells cultured with high glucose in vitro were analyzed by gene chip. To analyze its role in the occurrence and development of diabetic retinopathy. Methods Diabetic SD rats were induced by intraperitoneal injection of streptozotocin (STZ) (65 mg/kg) and control rats were treated with the same dose of sodium citrate solution. The blood glucose was measured by rat tail blood collection at 72 hours after injection. After 7 days blood glucose 鈮，

本文編號(hào)：2299039