本文選題：干擾素誘導(dǎo)蛋白-10 + 角膜堿燒傷��； 參考：《蘇州大學(xué)》2012年碩士論文

【摘要】：背景與目的 角膜作為重要的屈光介質(zhì)，生理情況下透明、無血管，但在炎癥、外傷、缺氧和水腫等病理狀態(tài)下，保持角膜無血管狀態(tài)的促血管形成因子與抑血管形成因子之間的平衡被打破，周圍毛細(xì)血管就會(huì)由角膜緣血管網(wǎng)侵入角膜，形成角膜新生血管（cornealneovascularization, CNV），從而導(dǎo)致視力減退甚至失明。干擾素誘導(dǎo)蛋白-10（interferon-inducible protein-10, IP-10）可作為趨化因子介導(dǎo)Th1型炎癥反應(yīng)，也可作為血管新生抑制劑抑制血管生成，抑制腫瘤生長(zhǎng)和轉(zhuǎn)移，參與血管重塑等過程。有關(guān)IP-10在CNV中的作用國內(nèi)外尚未見報(bào)道。本實(shí)驗(yàn)通過外源性小鼠IP-10蛋白對(duì)堿燒傷后新生血管形成的不同階段進(jìn)行局部干預(yù)，初步探討其對(duì)CNV形成的作用及機(jī)制，以期對(duì)新生血管性角膜疾病的研究提供理論依據(jù)及治療靶點(diǎn)。 材料與方法 1、采用浸潤(rùn)1mol/L NaOH的2×2mm濾紙貼附左眼角膜中央40s的方法制作小鼠角膜堿燒傷誘導(dǎo)CNV模型，角膜堿燒傷后1d及7d開始每日三次局部滴用IP-10蛋白10mg/L共7d分別作為早期實(shí)驗(yàn)組（10眼）和中晚期實(shí)驗(yàn)組（5眼），，各自的對(duì)照組均予質(zhì)量分?jǐn)?shù)0.2%透明質(zhì)酸鈉點(diǎn)眼（分別為11眼和6眼）。早期干預(yù)組于造模后2w，中晚期干預(yù)組于造模后3w分離角膜組織，以CD31免疫熒光標(biāo)記法標(biāo)記新生血管，比較對(duì)照組及實(shí)驗(yàn)組CNV面積。 2、收集早期干預(yù)第2d及第4d小鼠的角膜組織，用RT-PCR法檢測(cè)并比較各組小鼠角膜組織中趨化因子受體3（chemokine receptor3, CXCR3）、血管內(nèi)皮生長(zhǎng)因子（vascular endothelial growth factor, VEGF）及轉(zhuǎn)化生長(zhǎng)因子-β1（transforminggrowth factor-β1, TGF-β1）mRNA的表達(dá)情況。 3、分離早期干預(yù)第2d及第4d小鼠的角膜組織，制作鋪片，免疫組織化學(xué)法以F4/80標(biāo)記巨噬細(xì)胞，比較其在角膜組織中的浸潤(rùn)情況。 結(jié)果 1、早期干預(yù)對(duì)照組小鼠CNV占角膜總面積的比例為（88.67±3.23）%，IP-10實(shí)驗(yàn)組小鼠CNV面積為（70.06±3.68）%，兩組比較差異有統(tǒng)計(jì)學(xué)意義（t=3.77，P=0.0013）。中晚期干預(yù)對(duì)照組小鼠CNV占角膜總面積的比例為（87.33±5.50）%，IP-10實(shí)驗(yàn)組CNV面積為（86.56±5.58）%，兩組比較差異無統(tǒng)計(jì)學(xué)意義（t=1.26，P0.05）。 2、 IP-10早期干預(yù)第2d及第4d的小鼠角膜組織中CXCR3表達(dá)較對(duì)照組同期值明顯升高（t=3.13、3.07，P0.05），VEGF表達(dá)降低（t=5.99、6.27，P0.01），TGF-β1表達(dá)降低（t=8.50，P0.01；t=4.53，P0.05）。 3、 IP-10早期干預(yù)第2d及第4d的小鼠角膜組織中巨噬細(xì)胞的浸潤(rùn)實(shí)驗(yàn)組與對(duì)照組相比差異無統(tǒng)計(jì)學(xué)意義（t=1.65，P0.05；t=0.59，P0.05）。 結(jié)論 1.角膜堿燒傷后外源性IP-10蛋白早期干預(yù)可抑制CNV的形成，中晚期干預(yù)對(duì)角膜堿燒傷誘導(dǎo)的CNV無明顯作用； 2.角膜堿燒傷后外源性IP-10蛋白早期干預(yù)可通過上調(diào)CXCR3和下調(diào)VEGF及TGF-β1的表達(dá)抑制CNV的形成，但對(duì)巨噬細(xì)胞的浸潤(rùn)并無改變。
[Abstract]:Background and purpose The cornea, as an important refractive medium, is transparent and vascular free under physiological conditions, but in pathological conditions such as inflammation, trauma, hypoxia and edema. When the balance between angiogenic factors and angiogenic factors, which maintain the anangiogenic state of the cornea, is broken, the surrounding capillaries will invade the cornea by the limbal vascular network, forming the corneal neovascularization, CNV, which leads to the loss of vision and even blindness. Interferon-inducible protein-10 (IP-10) can act as a chemokine to mediate Th1 type inflammation and as a angiogenic inhibitor to inhibit angiogenesis, inhibit tumor growth and metastasis, and participate in vascular remodeling. The role of IP-10 in CNV has not been reported at home and abroad. In this experiment, exogenous mouse IP-10 protein was used to intervene in different stages of neovascularization after alkali burn, and to explore its effect on CNV formation and its mechanism. In order to provide theoretical basis and therapeutic targets for the study of neovascular corneal diseases. Materials and methods 1. The CNV model was induced by alkali burn of mouse cornea by applying 2 脳 2mm filter paper of infiltrating 1mol/L NaOH to the center of left cornea for 40 s. 10 eyes of the early experimental group and 5 eyes of the middle and late stage group were treated with IP-10 protein 10mg/L three times a day from 1 d and 7 d after alkali burn. The control groups were given 0.2% sodium hyaluronate (11 eyes and 6 eyes) respectively. Corneal tissue was isolated from the early intervention group at 2 weeks after modeling and the middle and late intervention group at 3 weeks after modeling. The neovascularization was labeled with CD31 immunofluorescence labeling method. The area of CNV in the control group and experimental group was compared with that in the control group and experimental group. 2. The corneal tissues of mice at the 2nd and 4th day after early intervention were collected. The expression of chemokine receptor 3, CXCR3, vascular endothelial growth factor (endothelial growth factor, VEGF) and transforming growth factor-尾 1(transforminggrowth factor- 尾 1, TGF- 尾 1)mRNA were detected and compared by RT-PCR method. 3. The cornea tissues of mice treated with early intervention on day 2 and day 4 were separated and prepared. Macrophages were labeled with F4 / 80 by immunohistochemical method to compare the infiltration of macrophages in cornea tissue. Result 1. The ratio of CNV to total corneal area in the early intervention control group was 88.67 鹵3.23 and the area of CNV in the IP-10 group was 70.06 鹵3.68. The difference between the two groups was statistically significant. The ratio of CNV to the total corneal area in the control group was 87.33 鹵5.50 and the area of CNV in the IP-10 experimental group was 86.56 鹵5.58. There was no significant difference between the two groups. 2. Compared with the control group, the expression of CXCR3 in the cornea of mice treated with IP-10 on the 2nd and 4th day was significantly higher than that of the control group. The expression of TGF- 尾 1 in the cornea of mice treated with IP-10 at the early stage was significantly higher than that in the control group (P 0.05). 3. There was no significant difference between the experimental group and the control group in the infiltration of macrophages in the cornea of the mice on the 2nd and 4th day after early intervention by IP-10. There was no significant difference between the experimental group and the control group. There was no significant difference between the experimental group and the control group. Conclusion 1. The early intervention of exogenous IP-10 protein after alkali burn could inhibit the formation of CNV, but the late intervention had no obvious effect on CNV induced by alkali burn of cornea. 2. The early intervention of exogenous IP-10 protein after alkali burn could inhibit the formation of CNV by up-regulating CXCR3 and down-regulating the expression of VEGF and TGF- 尾 1, but did not change the infiltration of macrophages.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R772.2

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