多波長激光誘導(dǎo)豚鼠形覺剝奪性高度近視脈絡(luò)膜新生血管模型建立的研究
發(fā)布時間：2018-12-17 16:45
【摘要】：目的：探討多波長激光誘導(dǎo)的形覺剝奪性高度近視豚鼠脈絡(luò)膜新生血管模型建立的可行性。 方法：1.多波長激光擊破Bruch’s膜：3周大雄性雙眼屈光參差大于6.00D三色豚鼠9只（18眼）分為激光后7d,14d,28d三組，雙眼A-Scan測量眼軸，取每只鼠近視屈光度更高眼作為實(shí)驗(yàn)眼，另一側(cè)眼為對照眼。采用多波長激光（532nm）擊破Bruch’s膜對豚鼠雙眼行視網(wǎng)膜光凝，激光功率、光斑直徑、曝光時間分別為400mW、50μm、0.1s，每只眼在視盤上方行8個光凝斑點(diǎn)。于光凝后三個時間點(diǎn)行眼底彩照和吲哚青綠血管造影（indocyanine green angiography, ICGA），檢查后處死豚鼠，摘取眼球制作標(biāo)本，進(jìn)行脈絡(luò)膜鋪片及組織病理學(xué)觀察。 2.多波長激光不擊破Bruch’s膜：3周大雄性雙眼屈光參差小于2.00D三色豚鼠30只，在右眼面罩法形覺剝奪誘導(dǎo)近視6周后，多波長激光（532nm）不擊破Bruch’s膜雙眼光凝，分為光凝后7d,14d,28d三組，激光功率為120mW，直徑200μm，曝光時間為0.1s，每只眼在視盤上方行10個光凝斑點(diǎn)。各組分別于光凝后三個時間點(diǎn)行眼底彩照和ICGA，檢查后處死豚鼠，摘取眼球制作標(biāo)本，進(jìn)行脈絡(luò)膜鋪片及組織病理學(xué)觀察，并通過免疫組化等分子生物學(xué)方法分析VEGF，MMP-2等因子在實(shí)驗(yàn)眼和對照眼視網(wǎng)膜等組織的蛋白表達(dá)情況。 結(jié)果：1.光凝后7d出現(xiàn)CNV，14d達(dá)高峰，28d出現(xiàn)瘢痕化。以ICGA激光斑熒光充盈為準(zhǔn)，14天時造模成功率為66.7%，且此時光鏡下可見視網(wǎng)膜下CNV形成，脈絡(luò)膜鋪片可見激光斑邊緣呈網(wǎng)簇狀高熒光斑，半定量分析三個時間點(diǎn)CNV面積，實(shí)驗(yàn)眼CNV面積均大于對照眼，且14d時，實(shí)驗(yàn)眼CNV面積最大，28d時，，雙眼CNV面積均減小。 2.面罩法形覺剝奪4周后誘導(dǎo)出-6.00D的近視，戴面罩眼眼軸較對照組明顯增長，且主要是玻璃體腔延長。激光光凝后，ICGA三個時間點(diǎn)激光斑均可見；脈絡(luò)膜鋪片和光鏡下均可見在視網(wǎng)膜下7d出現(xiàn)CNV，14dCNV達(dá)高峰，28d出現(xiàn)瘢痕化；免疫組化結(jié)果顯示形覺剝奪眼VEGF在脈絡(luò)膜毛細(xì)血管處高表達(dá)，MMP-2在視網(wǎng)膜色素上皮層處表達(dá)更多。 結(jié)論：多波長激光擊破Bruch’s膜可以誘導(dǎo)豚鼠CNV，并且屈光度高的豚鼠實(shí)驗(yàn)眼CNV面積比屈光度低的對照眼大，可能是由于屈光度高的實(shí)驗(yàn)眼眼軸增長導(dǎo)致視網(wǎng)膜變薄，從而為新生血管生長提供了更多的缺血缺氧等條件。多波長激光不擊破Bruch’s膜可以在已經(jīng)誘導(dǎo)出高度近視的豚鼠眼上誘導(dǎo)出CNV，VEGF和MMP-2參與了高度近視CNV的形成過程。這與人類病理性近視CNV發(fā)病機(jī)制較為相似，該模型可用于病理性近視CNV病因?qū)W、發(fā)病機(jī)制及藥物治療效果的研究。
[Abstract]:Objective: to investigate the feasibility of multiwavelength laser induced choroidal neovascularization model in guinea pigs with high myopia of form deprivation. Method 1: 1. Bruch's film was broken by multi-wavelength laser: 9 male anisometropia > 6.00D (18 eyes) were divided into three groups (7 days, 14 days and 28 days after laser). The axial axis of the eyes was measured by A-Scan. Each rat myopia diopter higher eye as experimental eye, the other side of the eye as a control eye. The retinal photocoagulation, laser power, spot diameter, and exposure time of the Bruch's film were observed in guinea pig eyes by using a multi-wavelength laser (532nm) method. The laser power, spot diameter and exposure time were 400mW ~ (50) 渭 m ~ (-1) ~ (0.1) s, respectively. Eight photocoagulation spots were performed in each eye over the optic disc. The guinea pigs were sacrificed at three time points after photocoagulation by fundus photography and indole green angiography (indocyanine green angiography, ICGA),). The eyeball specimens were removed and the choroidal smears and histopathology were observed. 2. The multiwavelength laser did not break the Bruch's film: the anisometropia of 3 weeks old male eyes was less than 2.00D in 30 guinea pigs. After the myopia was induced by right eye mask form deprivation for 6 weeks, the multiwavelength laser (532nm) did not break the Bruch's film binocular photocoagulation. They were divided into three groups: laser power 120 MW, diameter 200 渭 m, exposure time 0.1 s. Each eye was given 10 spots above the optic disc. Three time points after photocoagulation, the guinea pigs were killed by fundus colorimetry and ICGA, examination respectively. The eyeballs were taken out to make specimens, choroidal smears and histopathology were observed, and VEGF, was analyzed by immunohistochemistry and other molecular biological methods. Expression of MMP-2 and other factors in retinal tissues of experimental eyes and control eyes. Results: 1. CNV,14d peaked at 7 days after photocoagulation, and scarring occurred on 28 days after photocoagulation. On the basis of ICGA laser spot fluorescence filling, the successful rate of modeling was 66.7% at 14 days, and the formation of subretinal CNV was observed under light microscope, and the laser spot edge of choroidal patch was shown as a reticular high fluorescence spot. The area of CNV was semi-quantitatively analyzed at three time points. The CNV area of the experimental eyes was larger than that of the control eyes, and the CNV area of the experimental eyes was the largest at 14 days, and the CNV area of both eyes decreased at 28 days. 2. After 4 weeks of face mask deprivation, -6.00D myopia was induced. The ocular axis in mask was significantly larger than that in the control group, and the vitreous cavity was lengthened. After laser photocoagulation, laser spots were observed at three time points of ICGA, and CNV,14dCNV reached the peak at 7 days under retina and scarred at 28 days. Immunohistochemical results showed that VEGF was highly expressed in choroidal capillaries in form-deprived eyes, and MMP-2 was more expressed in retinal pigment epithelium. Conclusion: the area of CNV in guinea pig experimental eyes with high diopter is larger than that in experimental eyes with low diopter, which may be due to the retinal thinning caused by the axial growth of experimental eyes with high diopter. This provides more conditions for neovascularization, such as ischemia and hypoxia. Multiwavelength laser without breaking the Bruch's membrane can induce CNV,VEGF and MMP-2 to participate in the formation of CNV in the eyes of guinea pigs that have already induced high myopia. This model is similar to the pathogenesis of human pathological myopia (CNV). This model can be used to study the etiology, pathogenesis and therapeutic effect of CNV in pathological myopia.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R778.11
本文編號：2384466
[Abstract]:Objective: to investigate the feasibility of multiwavelength laser induced choroidal neovascularization model in guinea pigs with high myopia of form deprivation. Method 1: 1. Bruch's film was broken by multi-wavelength laser: 9 male anisometropia > 6.00D (18 eyes) were divided into three groups (7 days, 14 days and 28 days after laser). The axial axis of the eyes was measured by A-Scan. Each rat myopia diopter higher eye as experimental eye, the other side of the eye as a control eye. The retinal photocoagulation, laser power, spot diameter, and exposure time of the Bruch's film were observed in guinea pig eyes by using a multi-wavelength laser (532nm) method. The laser power, spot diameter and exposure time were 400mW ~ (50) 渭 m ~ (-1) ~ (0.1) s, respectively. Eight photocoagulation spots were performed in each eye over the optic disc. The guinea pigs were sacrificed at three time points after photocoagulation by fundus photography and indole green angiography (indocyanine green angiography, ICGA),). The eyeball specimens were removed and the choroidal smears and histopathology were observed. 2. The multiwavelength laser did not break the Bruch's film: the anisometropia of 3 weeks old male eyes was less than 2.00D in 30 guinea pigs. After the myopia was induced by right eye mask form deprivation for 6 weeks, the multiwavelength laser (532nm) did not break the Bruch's film binocular photocoagulation. They were divided into three groups: laser power 120 MW, diameter 200 渭 m, exposure time 0.1 s. Each eye was given 10 spots above the optic disc. Three time points after photocoagulation, the guinea pigs were killed by fundus colorimetry and ICGA, examination respectively. The eyeballs were taken out to make specimens, choroidal smears and histopathology were observed, and VEGF, was analyzed by immunohistochemistry and other molecular biological methods. Expression of MMP-2 and other factors in retinal tissues of experimental eyes and control eyes. Results: 1. CNV,14d peaked at 7 days after photocoagulation, and scarring occurred on 28 days after photocoagulation. On the basis of ICGA laser spot fluorescence filling, the successful rate of modeling was 66.7% at 14 days, and the formation of subretinal CNV was observed under light microscope, and the laser spot edge of choroidal patch was shown as a reticular high fluorescence spot. The area of CNV was semi-quantitatively analyzed at three time points. The CNV area of the experimental eyes was larger than that of the control eyes, and the CNV area of the experimental eyes was the largest at 14 days, and the CNV area of both eyes decreased at 28 days. 2. After 4 weeks of face mask deprivation, -6.00D myopia was induced. The ocular axis in mask was significantly larger than that in the control group, and the vitreous cavity was lengthened. After laser photocoagulation, laser spots were observed at three time points of ICGA, and CNV,14dCNV reached the peak at 7 days under retina and scarred at 28 days. Immunohistochemical results showed that VEGF was highly expressed in choroidal capillaries in form-deprived eyes, and MMP-2 was more expressed in retinal pigment epithelium. Conclusion: the area of CNV in guinea pig experimental eyes with high diopter is larger than that in experimental eyes with low diopter, which may be due to the retinal thinning caused by the axial growth of experimental eyes with high diopter. This provides more conditions for neovascularization, such as ischemia and hypoxia. Multiwavelength laser without breaking the Bruch's membrane can induce CNV,VEGF and MMP-2 to participate in the formation of CNV in the eyes of guinea pigs that have already induced high myopia. This model is similar to the pathogenesis of human pathological myopia (CNV). This model can be used to study the etiology, pathogenesis and therapeutic effect of CNV in pathological myopia.
【學(xué)位授予單位】：青海大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R778.11
【參考文獻(xiàn)】
相關(guān)期刊論文 前5條
1 楊蓓;劉桂香;;豚鼠短期形覺剝奪性近視屈光度眼軸及鞏膜改變[J];國際眼科雜志;2009年10期
2 楊秀梅;王雨生;;脈絡(luò)膜新生血管的動物模型[J];國際眼科縱覽;2006年03期
3 曾軍,姜德詠,劉湘平,朱曉華,唐羅生;MMP-2和MMP-9在脈絡(luò)膜新生血管膜的表達(dá)[J];眼科新進(jìn)展;2004年06期
4 趙世紅,何守志;氪激光誘導(dǎo)的大鼠脈絡(luò)膜新生血管模型研究[J];中華眼科雜志;2003年05期
5 歐陽朝祜,褚仁遠(yuǎn),胡文政;哌侖西平對豚鼠透鏡誘導(dǎo)性近視眼的作用[J];中華眼科雜志;2003年06期
本文編號：2384466
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