【摘要】： [目的]：研究氧誘導(dǎo)的視網(wǎng)膜新生血管形成的機(jī)制及防治方法。 [方法]：20只新生C57BL/6J小鼠隨機(jī)分為對照組和實(shí)驗(yàn)組,各10只。實(shí)驗(yàn)組P7置75%氧濃度環(huán)境中,P12返回正�？諝庵�,對照組始終置于正常空氣環(huán)境中,P17行熒光素灌注造影、視網(wǎng)膜鋪片及病理切片,觀察視網(wǎng)膜新生血管生成情況；幼鼠P12、P14、P17對照組和實(shí)驗(yàn)組各6只,取視網(wǎng)膜,應(yīng)用realtime-PCR定量檢測各時(shí)間點(diǎn)Annexin A2mRNA和TPAmRNA水平；HSS治療組10只OIR模型小鼠腹腔注射飽和氫鹽水、HSS對照組10只OIR模型小鼠腹腔注射生理鹽水,正常組10只小鼠分別于P17用高分子量熒光素灌注造影觀察血管分布與形態(tài),HE染色計(jì)數(shù)突破內(nèi)界膜的血管內(nèi)皮細(xì)胞核數(shù),用realtime-PCR和免疫組化檢測視網(wǎng)膜VEGF表達(dá),并測定丙二醛含量。 [結(jié)果]：1.熒光素灌注造影結(jié)果顯示實(shí)驗(yàn)組小鼠視網(wǎng)膜中央呈無灌注區(qū)域,視網(wǎng)膜血管迂曲擴(kuò)張、熒光素滲漏,對照組小鼠視網(wǎng)膜血管分布均勻,未見無灌注區(qū)和熒光素滲漏；病理切片可見實(shí)驗(yàn)組小鼠突破內(nèi)界膜的血管內(nèi)皮細(xì)胞核數(shù)為79.70±7.57,對照組為0.28±0.12,兩組具有顯著差異(P0.01)。2.P12和P17時(shí),實(shí)驗(yàn)組與對照組Annexin A2mRNA和TPA mRNA的表達(dá)量無顯著差異(P0.05)；P14時(shí),實(shí)驗(yàn)組Annexin A2mRNA和TPA mRNA表達(dá)量均比對照組高,且具有顯著差異(P0.01)。3.HSS治療組熒光素灌注造影未見無灌注區(qū)、新生血管叢及熒光素滲漏；HSS對照組熒光素灌注造影視網(wǎng)膜視乳頭周邊可見大片無灌注區(qū),視網(wǎng)膜血管不規(guī)則擴(kuò)、迂曲,無灌注區(qū)周圍可見大量新生血管叢,伴明顯熒光素滲漏；正常組熒光素灌注造影視網(wǎng)膜未見無灌注區(qū)及熒光素滲漏。病理切片結(jié)果顯示HSS治療組、HSS對照組和正常組突破內(nèi)界膜的血管內(nèi)皮細(xì)胞核數(shù)分別為41.00±8.01、79.70±7.57和0.90±1.28,HSS治療組與正常組無統(tǒng)計(jì)學(xué)差異(P0.05),HSS治療組與HSS對照組具有顯著差異(P0.01)；免疫組織化學(xué)實(shí)驗(yàn)顯示VEGF表達(dá)于神經(jīng)節(jié)細(xì)胞層、內(nèi)核層和色素上皮細(xì)胞層,VEGF蛋白陽性表達(dá)HSS治療組、HSS對照組和正常組分別為1.46±0.01、2.92±0.70和1.30±0.06；HSS治療組、HSS對照組和正常組VEGFmRNA表達(dá)量分別為1.94±0.12、7.40±0.04和1.00±0.03,HSS治療組與正常組之間無統(tǒng)計(jì)學(xué)意義(P0.05),HSS治療組與HSS對照組有顯著性差異(P0.01)；小鼠視網(wǎng)膜中MDA含量HSS治療組為16.07±1.05 nmol/mg prot, HSS對照組為22.42±2.24 nmol/mg prot,正常組為5.17±4.23 nmol/mg prot, HSS治療組與HSS對照組具有顯著差異(P0.01),正常組與HSS對照組有顯著差別(P0.01)。 [結(jié)論]：氧誘導(dǎo)視網(wǎng)膜新生血管小鼠模型穩(wěn)定、可靠、重復(fù)性高,可做為研究視網(wǎng)膜新生血管疾病發(fā)病機(jī)制和防治方法的動物模型；Annexin A2和TPA與氧誘導(dǎo)視網(wǎng)膜新生血管的形成密切相關(guān)；飽和氫鹽水對氧誘導(dǎo)的視網(wǎng)膜新生血管的形成有抑制作用。
[Abstract]:Objective: to study the mechanism, prevention and treatment of oxygen-induced retinal neovascularization. [methods]: 20 newborn C57BL/6J mice were randomly divided into control group (n = 10) and experimental group (n = 10). P7 was placed in 75% oxygen concentration in the experimental group, P12 returned to the normal air, and the control group was always in the normal air. P17 was performed fluorescein perfusion angiography, retinal preparation and pathological sections to observe the retinal neovascularization. The retina of P12, P14, P17 control group and experimental group were collected and the levels of Annexin A2mRNA and TPAmRNA were measured by realtime-PCR. In the HSS treatment group, 10 OIR mice were injected with saturated hydrogen saline intraperitoneally, and 10 OIR mice in the HSS control group were injected with saline intraperitoneally. In the normal group, 10 mice in the normal group were given high molecular weight fluorescein perfusion angiography to observe the distribution and morphology of the blood vessels. HE staining was used to count the nuclei of vascular endothelial cells which broke through the inner limiting membrane. The expression of VEGF in retina was detected by realtime-PCR and immunohistochemistry, and the content of malondialdehyde (MDA) was measured. [results]: 1. The results of fluorescein perfusion angiography showed that there was no perfusion area in the central retina of the experimental group, the retinal vessels were dilated roundly and fluorescein leakage was found in the experimental group, while the retinal vessels in the control group were evenly distributed, no perfusion area and no fluorescein leakage were found. Pathological sections showed that the number of endothelial cell nuclei breaking through the inner limiting membrane was 79.70 鹵7.57 in the experimental group and 0.28 鹵0.12 in the control group. There was significant difference between the two groups (P0.01). 2. P12 and P17 were significantly different from those in the control group (P < 0.01). There was no significant difference in the expression of Annexin A2mRNA and TPA mRNA between the experimental group and the control group (P0.05). At P14, the expression of Annexin A2mRNA and TPA mRNA in experimental group was higher than that in control group (P0.01). 3. There was no perfusion area, neovascularization and fluorescein leakage in fluorescein perfusion angiography in HSS group. In the HSS control group, large areas of no perfusion were seen around the optic papillae by fluorescein perfusion. The retinal vessels were dilated irregularly, and a large number of neovascularization and fluorescein leakage were seen around the no perfusion areas. In the normal group, no fluorescein leakage and no perfusion were found in the omentum. Pathological sections showed that the number of endothelial cell nuclei breaking through the inner boundary membrane in HSS treatment group, HSS control group and normal group were 41.00 鹵8.01, 79.70 鹵7.57 and 0.90 鹵1.28, respectively. There was no significant difference between HSS treatment group and normal group (P0.05). There was significant difference between HSS treatment group and HSS control group (P0.01). Immunohistochemistry showed that VEGF was expressed in ganglion cell layer, inner nuclear layer and pigment epithelial layer. The positive expression of VEGF protein in HSS group, HSS control group and normal group were 1.46 鹵0.01,2.92 鹵0.70 and 1.30 鹵0.06, respectively. The expression of VEGFmRNA in HSS treatment group, HSS control group and normal group was 1.94 鹵0.12, 7.40 鹵0.04 and 1.00 鹵0.03, respectively. There was no significant difference between the two groups (P0.05). There was significant difference between HSS treatment group and HSS control group (P0.01). The content of MDA in the retina of mice was 16.07 鹵1.05 nmol/mg prot, HSS in HSS group, 22.42 鹵2.24 nmol/mg prot, in control group, 5.17 鹵4.23 nmol/mg prot, HSS in normal group and 5.17 鹵4.23 nmol/mg prot, HSS in HSS control group (P0.01). There was significant difference between normal group and HSS control group (P0.01). [conclusion]: oxygen-induced retinal neovascularization mouse model is stable, reliable and reproducible. It can be used as an animal model to study the pathogenesis, prevention and treatment of retinal neovascularization disease. Annexin A2 and TPA are closely related to the formation of retinal neovascularization induced by oxygen, and saturated hydrogen salt can inhibit the formation of oxygen-induced retinal neovascularization.
【學(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2010
【分類號】：R774.1

【共引文獻(xiàn)】

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2 王欣東;丁苯酞對血管性癡呆大鼠學(xué)習(xí)記憶障礙的實(shí)驗(yàn)研究[D];鄭州大學(xué);2011年

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10 孫漢勇;氫鹽水對小鼠實(shí)驗(yàn)性肝損傷保護(hù)作用及其機(jī)制研究[D];第二軍醫(yī)大學(xué);2010年

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