本文選題：內(nèi)質(zhì)網(wǎng)相關(guān)降解 + 糖尿病視網(wǎng)膜病變��； 參考：《華中科技大學(xué)》2011年碩士論文

【摘要】：目的:內(nèi)質(zhì)網(wǎng)相關(guān)降解(ER-associated degradation, ERAD)是內(nèi)質(zhì)網(wǎng)中重要的蛋白質(zhì)質(zhì)量監(jiān)控系統(tǒng),能夠降解錯(cuò)誤折疊的蛋白,避免細(xì)胞發(fā)生內(nèi)質(zhì)網(wǎng)應(yīng)激,內(nèi)質(zhì)網(wǎng)應(yīng)激與糖尿病視網(wǎng)膜病變的發(fā)生、發(fā)展有密切關(guān)系。本研究通過建立SD大鼠糖尿病模型,檢測(cè)ERAD作用途徑相關(guān)因子在糖尿病鼠視網(wǎng)膜中的表達(dá)變化,從而探討ERAD相關(guān)因子在糖尿病視網(wǎng)膜病變的早期階段的發(fā)生和發(fā)展中可能存在的作用。 方法:用大劑量鏈鏈脲佐菌素(Streptozotocin ,STZ)一次性腹腔注射造成糖尿病SD大鼠模型,以同齡SD大鼠腹腔注射等量的檸檬酸鈉緩沖液為對(duì)照組。自行設(shè)計(jì)實(shí)時(shí)定量PCR芯片檢測(cè)內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)因子在一月糖尿病鼠視網(wǎng)膜中的表達(dá)變化,Western blot、免疫組化檢測(cè)一月糖尿病鼠視網(wǎng)膜中ERAD相關(guān)因子Herp蛋白的表達(dá)變化。體外培養(yǎng)人視網(wǎng)膜微血管內(nèi)皮細(xì)胞,高糖干預(yù)后,免疫熒光檢測(cè)Herp蛋白的表達(dá)變化。 結(jié)果:實(shí)時(shí)定量PCR芯片結(jié)果顯示:一月糖尿病鼠視網(wǎng)膜中有13個(gè)內(nèi)質(zhì)網(wǎng)應(yīng)激相關(guān)因子的mRNA水平明顯下調(diào)(P0.05),涵蓋內(nèi)質(zhì)網(wǎng)應(yīng)激中了8個(gè)作用途徑,其中ERAD相關(guān)因子Herp, NUC, Os9, ADO15的表達(dá)明顯下調(diào)(P0.05)。Western blot與免疫熒光提示,Herp蛋白在一月糖尿病模型鼠視網(wǎng)膜中的表達(dá)明顯下調(diào)(P0.05);體外培養(yǎng)人視網(wǎng)膜微血管內(nèi)皮細(xì)胞在高糖干預(yù)后,免疫熒光顯示Herp在蛋白水平的表達(dá)明顯下調(diào)(P0.05)。 結(jié)論:內(nèi)質(zhì)網(wǎng)應(yīng)激與糖尿病視網(wǎng)膜病變的發(fā)生與發(fā)展有密切的關(guān)系, ERAD作用途徑參與了糖尿病視網(wǎng)膜病變的發(fā)生和發(fā)展,Herp在該過程中發(fā)揮了重要的的作用。
[Abstract]:Objective: ER-associated degradation (ERAD) is an important protein quality monitoring system in the endoplasmic reticulum (ER), which can degrade misfolded proteins and avoid endoplasmic reticulum stress, endoplasmic reticulum stress and diabetic retinopathy. Development is closely related. In this study, SD rat model of diabetes mellitus was established to detect the expression of ERAD pathway related factors in the retina of diabetic rats. To explore the role of ERAD related factors in the early stage of diabetic retinopathy. Methods: a diabetic SD rat model was established by intraperitoneal injection of streptozotocin (STZ) in a single dose, and sodium citrate buffer was injected intraperitoneally to the same age SD rats as control group. A real-time quantitative PCR chip was designed to detect the expression of ER stress-related factors in the retina of diabetic rats in January. The expression of ERAD-related factor Herp protein in the retina of diabetic rats in January was detected by immunohistochemistry. Human retinal microvascular endothelial cells were cultured in vitro. The expression of Herp protein was detected by immunofluorescence after high glucose intervention. Results: the results of real-time quantitative PCR microarray showed that the mRNA levels of 13 ER stress-related factors in the retina of diabetic rats were significantly down-regulated (P0.05), covering 8 pathways of endoplasmic reticulum stress. The expression of Herp, NUC, Os9, ADO15 was significantly down-regulated (P0.05). Western blot and immunofluorescence indicated that Herp protein expression was significantly down-regulated in the retina of diabetic rats in January (P0.05); in vitro cultured human retinal microvascular endothelial cells (RMECs) were treated with high glucose. Immunofluorescence showed that Herp expression was significantly down-regulated at protein level (P0.05). Conclusion: endoplasmic reticulum stress is closely related to the occurrence and development of diabetic retinopathy. ERAD plays an important role in the pathogenesis and development of diabetic retinopathy.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2011
【分類號(hào)】：R774.1

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