本文選題：胰島素樣生長(zhǎng)因子-1 + 糖尿病��； 參考：《山東大學(xué)》2010年碩士論文

【摘要】： 目的糖尿病角膜創(chuàng)傷愈合延遲是目前眼科臨床最常見問題之一,治療棘手,發(fā)病機(jī)制尚不明確。IGF-1可促進(jìn)細(xì)胞遷移、增殖,抑制細(xì)胞凋亡,在創(chuàng)傷愈合中發(fā)揮重要作用。本實(shí)驗(yàn)通過建立糖尿病大鼠角膜創(chuàng)傷模型,檢測(cè)創(chuàng)傷愈合過程中IGF-1的表達(dá),從分子水平探求糖尿病角膜創(chuàng)傷愈合延遲的可能機(jī)制。 方法1.建立糖尿病大鼠模型：65只雄性Wistar大鼠,體重170-200g,隨機(jī)分為糖尿病組和正常對(duì)照組。糖尿病組腹腔注射鏈脲霉素(STZ)制作糖尿病大鼠模型(n=35),正常對(duì)照組腹腔注射等量檸檬酸鹽緩沖液(n=30)。造模后飼養(yǎng)8周,每周測(cè)血糖和體重。2.建立大鼠角膜創(chuàng)傷模型：用直徑5.0毫米的環(huán)鉆在角膜中央標(biāo)記創(chuàng)傷區(qū),刮除標(biāo)記區(qū)內(nèi)的角膜上皮。3.分別在角膜上皮移除術(shù)后不同時(shí)間點(diǎn)(0h,12h,24h,36h,48h,60h,72h,4d-16d),在裂隙燈下觀察角膜創(chuàng)傷愈合及感染情況；進(jìn)行熒光素鈉染色實(shí)驗(yàn),拍照記錄并分析角膜上皮創(chuàng)傷愈合率。4.分別在角膜上皮移除術(shù)后不同時(shí)間點(diǎn)(0h,12h,1d,2d,3d,4d,5d,7d)處死正常大鼠(n=24)及糖尿病大鼠(n=24),取角膜組織,HE染色分析角膜創(chuàng)傷愈合的病理組織學(xué)變化；real time RT-PCR檢測(cè)IGF-1mRNA在不同時(shí)間點(diǎn)的表達(dá)；免疫熒光法檢測(cè)IGF-1的表達(dá)位置,分析累積光密度(integrated optical density, IOD)檢測(cè)IGF-1蛋白表達(dá)量。 結(jié)果1.糖尿病大鼠出現(xiàn)典型的“三多一少”癥狀：多飲、多食、多尿、消瘦,體重顯著低于正常大鼠(P0.01),血糖濃度持續(xù)顯著高于正常大鼠(P0.01)。到第8周時(shí),糖尿病大鼠體重比正常大鼠體重低25.8%(262.11±12.11g,353.44±10.85g,P0.01),血糖濃度比正常大鼠血糖濃度高4.63倍(18.43±1.04 mmol／L,3.98±0.78mmol／L,P0.01)。2.與正常大鼠相比,糖尿病大鼠角膜上皮創(chuàng)傷愈合速率顯著延遲。創(chuàng)傷后60h,正常大鼠角膜上皮創(chuàng)傷完全愈合,糖尿病大鼠角膜上皮創(chuàng)傷愈合率為80.7%,兩組的愈合率在24,36,48及60h均有顯著性差異(P0.01)。3.HE染色顯示,炎癥細(xì)胞在糖尿病大鼠角膜創(chuàng)傷區(qū)的出現(xiàn)時(shí)間延遲,停留時(shí)間延長(zhǎng),伴基質(zhì)水腫、大量角膜緣新生血管生成。4.未創(chuàng)傷時(shí),正常大鼠角膜中未檢測(cè)到IGF-1mRNA表達(dá),糖尿病大鼠角膜中可見少量表達(dá)。正常大鼠角膜中IGF-1 mRNA在創(chuàng)傷后1d表達(dá)上調(diào),2d達(dá)高峰,高峰持續(xù)至5d時(shí)表達(dá)下調(diào)。糖尿病大鼠角膜中IGF-1mRNA在創(chuàng)傷后2d表達(dá)上調(diào),3d達(dá)高峰,持續(xù)高表達(dá)。創(chuàng)傷后2d-4d,正常大鼠角膜中IGF-1mRNA的表達(dá)水平顯著高于糖尿病大鼠(P0.01),分別高4.8,1.6及2.2倍。5.未創(chuàng)傷時(shí),正常大鼠角膜中幾乎無IGF-1表達(dá),糖尿病大鼠角膜上皮及基質(zhì)中可見少量IGF-1表達(dá)。創(chuàng)傷后,正常大鼠創(chuàng)傷區(qū)角膜基質(zhì)及新覆蓋的角膜上皮中,IGF-1在傷后1d開始表達(dá),3d達(dá)高峰,5d表達(dá)量降低,但仍可見有表達(dá)。糖尿病大鼠角膜上皮及創(chuàng)傷區(qū)基質(zhì)中IGF-1在傷后1d上升,3d達(dá)高峰,5天時(shí)表達(dá)顯著下降。分析IOD結(jié)果顯示,正常大鼠角膜創(chuàng)傷愈合過程中IGF-1的表達(dá)量顯著高于糖尿病大鼠(P0.01),在1d,3d,5d時(shí),分別高4.6倍,2.2倍及2.4倍。 結(jié)論IGF-1表達(dá)延遲和表達(dá)不足可能是導(dǎo)致糖尿病角膜創(chuàng)傷愈合延遲的重要機(jī)制之一。
[Abstract]:Objective diabetic corneal wound healing delay is one of the most common clinical problems in Ophthalmology, the treatment is difficult. The pathogenesis is not clear.IGF-1 can promote cell migration, proliferation, inhibit cell apoptosis and play an important role in wound healing. This experiment was established by establishing a diabetic rat corneal wound model to detect IGF-1 in the healing process. To explore the possible mechanism of delayed healing of diabetic corneal injury from molecular level.
Method 1. the diabetic rat model was established: 65 male Wistar rats and weight 170-200g were randomly divided into diabetes group and normal control group. Diabetic rats were injected with streptozotocin (STZ) to make diabetic rat model (n=35). The normal control group was injected with equal amount of citrate buffer (n=30). After making the model for 8 weeks, the blood glucose and body were measured weekly. A rat model of corneal trauma was established with a 5 millimeter diameter ring with a diameter of 5 millimeters in the cornea. Corneal epithelium.3. was removed at different time points after removal of the corneal epithelium (0h, 12h, 24h, 36h, 48h, 60H, 72h, 4d-16d) in the corneal epithelium. The corneal wound healing and infection were observed under the slit lamp. Fluorescein sodium staining was performed under the slit lamp. Corneal epithelial wound healing rate (0h, 12h, 1D, 2D, 3D, 4D, 5D, 7D) were sacrificed to the normal rats (n=24) and diabetic rats (n=24) after the corneal epithelial removal, and the corneal tissue was taken to analyze the histopathological changes of corneal wound healing. The expression of IGF-1 was detected by immunofluorescence, and the expression of IGF-1 protein was analyzed by integrated optical density (IOD).
Results 1. diabetic rats had typical symptoms of "more than three more and one less": polydipsia, polyuria, emaciation, weight loss significantly lower than normal rats (P0.01), and the blood glucose concentration was significantly higher than that of normal rats (P0.01). By eighth weeks, the weight of diabetic rats was 25.8% (262.11 + 12.11g, 353.44 + 10.85g, P0.01), and the blood glucose concentration ratio was higher than that of normal rats. In normal rats, the blood glucose concentration was 4.63 times higher (18.43 + 1.04 mmol / L, 3.98 + 0.78mmol / L, P0.01).2., compared with normal rats, the healing rate of corneal epithelial wound in diabetic rats was significantly delayed. After trauma, the corneal epithelial wound healing was completely healed in 60H, and the healing rate of corneal epithelial wound in diabetic rats was 80.7%. The healing rate of the two groups was 24,36,4 The significant difference between 8 and 60H (P0.01).3.HE staining showed that the occurrence time of the inflammatory cells in the corneal wound area of the diabetic rats was delayed, the stay time was prolonged, the matrix edema and a large number of corneal limbus neovascularization were not wound, and the expression of IGF-1mRNA was not detected in the normal rat cornea, and a small amount of expression was seen in the cornea of diabetic rats. The expression of IGF-1 mRNA in the cornea of normal rats was up-regulated, 2D reached its peak, and the expression of IGF-1mRNA was down regulated when the peak was continued to 5D. The 2D expression of IGF-1mRNA in the cornea of diabetic rats was up to up, the peak of 3D reached its peak, and the expression of IGF-1mRNA in the cornea of normal rats was significantly higher than that of the diabetic rats (P0.01), and the high 4.8,1.6 was higher than that of the diabetic rats (P0.01). The high 4.8,1.6 in the cornea of normal rats was higher than that of the diabetic rats (P0.01). The 4.8,1.6 was higher than that of the diabetic rats. There was almost no IGF-1 expression in normal rats' cornea and a small amount of IGF-1 expression in the corneal epithelium and matrix of diabetic rats. After trauma, IGF-1 was expressed in 1D after injury in normal rats and in the newly covered corneal epithelium. The expression of 3D reached the peak and the expression of 5D decreased, but the expression of IGF-1 was still visible. The expression of diabetes was still visible. Diabetes was still expressed. The expression of diabetes was still obvious. The 1D increased after injury in the corneal epithelium and wound area of the rat, and the expression of 3D reached a peak at 5 days. The analysis of IOD showed that the expression of IGF-1 in the healing process of normal rats was significantly higher than that of diabetic rats (P0.01), which was 4.6 times, 2.2 times and 2.4 times higher in 1D, 3D, and 5D respectively.
Conclusion delayed expression of IGF-1 and insufficient expression may be one of the important mechanisms leading to delayed wound healing in diabetic patients.
【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R587.1;R772.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 李養(yǎng)軍,惠延年;IGF-I對(duì)視網(wǎng)膜新生血管形成的調(diào)控及其信號(hào)轉(zhuǎn)導(dǎo)機(jī)制[J];國際眼科雜志;2005年05期

2 張小玲;孫文濤;;糖尿病性視網(wǎng)膜病變患者房水中VEGF與IGF-1的定量測(cè)定及分析[J];國際眼科雜志;2008年05期

3 杜杰 ,艾靜 ,楊寶峰;原釩酸鈉對(duì)Ⅱ型糖尿病的降糖作用研究[J];哈爾濱醫(yī)科大學(xué)學(xué)報(bào);2003年04期

4 王萍,馮佑民;類胰島素生長(zhǎng)因子-Ⅰ和胰島素表現(xiàn)其功能的結(jié)構(gòu)和分子基礎(chǔ)[J];生物化學(xué)與生物物理進(jìn)展;1999年06期

5 付小兵,王亞平,孫同柱,楊銀輝;糖尿病慢性難愈合創(chuàng)面大鼠模型的制備[J];上海實(shí)驗(yàn)動(dòng)物科學(xué);1997年04期

6 張力,李樹濃;胰島素樣生長(zhǎng)因子-Ⅰ[J];國外醫(yī)學(xué)(生理、病理科學(xué)與臨床分冊(cè));1997年02期

7 馬波;惠延年;;胰島素樣生長(zhǎng)因子-1誘導(dǎo)糖尿病性白內(nèi)障晶狀體上皮細(xì)胞表達(dá)MCP-1[J];細(xì)胞與分子免疫學(xué)雜志;2007年08期

8 劉真喜,傅庭煥;IGF-1與細(xì)胞凋亡的研究進(jìn)展[J];醫(yī)學(xué)綜述;2001年10期

9 王正國;創(chuàng)傷修復(fù)的分子生物學(xué)研究[J];中華創(chuàng)傷雜志;2000年06期

10 許光武,俞茂華;胰島素樣生長(zhǎng)因子[J];中華內(nèi)分泌代謝雜志;2000年01期

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本文編號(hào)：2009339