【摘要】： 目的 青光眼是當(dāng)今世界范圍內(nèi)的主要致盲性眼病之一,由于眼壓超過(guò)視網(wǎng)膜節(jié)細(xì)胞(rentinal ganglion cells, RGCs)的承受范圍,引起RGCs凋亡造成視功能的進(jìn)行性不可逆性損害,但RGCs凋亡的機(jī)制尚不清楚。HSP是生物體細(xì)胞在不良環(huán)境因素作用下所產(chǎn)生的一組特殊的蛋白質(zhì),能增強(qiáng)細(xì)胞存活能力,保護(hù)細(xì)胞不受或少受傷害。其中熱休克蛋白70(heat shock protein, HSP70)是在中樞神經(jīng)系統(tǒng)含量最豐富的熱休克蛋白。早期研究證實(shí)抗?jié)兯幬锾嫫杖鹜?geranglgeranylacetone, GGA)可以明顯誘導(dǎo)胃腸粘膜、肝臟、心臟等組織的HSP70表達(dá)。新近研究發(fā)現(xiàn)一定劑量的替普瑞酮在中樞神經(jīng)系統(tǒng)也可以誘導(dǎo)HSP70表達(dá)。我們通過(guò)阻斷大鼠鞏膜淺層靜脈聯(lián)合絲裂霉素的方法阻斷房水的外引流途徑,使其眼壓升高,模擬慢性青光眼的發(fā)病過(guò)程。觀察替普瑞酮對(duì)大鼠慢性高眼壓視網(wǎng)膜HSP70表達(dá)的影響,為青光眼視神經(jīng)保護(hù)提供新的治療思路。 方法 1慢性高眼壓模型的制備 熱凝大鼠鞏膜淺層靜脈聯(lián)合應(yīng)用絲裂霉素阻斷房水外引流途徑制成模型；TONO-PEN XL型筆式眼壓計(jì)測(cè)量術(shù)前,術(shù)后即刻,1d,3d,7d,14d,21d,28d眼壓。 2 HE染色觀察視網(wǎng)膜形態(tài)學(xué)變化。 3正常視網(wǎng)膜,慢性高眼壓視網(wǎng)膜,慢性高眼壓+GGA后視網(wǎng)膜HSP70蛋白表達(dá)水平的初步研究。 應(yīng)用非生物素二步法免疫組化法檢測(cè)正常視網(wǎng)膜,慢性高眼壓視網(wǎng)膜,用藥后視網(wǎng)膜中HSP70蛋白表達(dá)的部位和程度。統(tǒng)計(jì)結(jié)果均以均數(shù)±標(biāo)準(zhǔn)差表示,組間比較應(yīng)用One-Way ANOVA分析,均用SPSS13.0軟件進(jìn)行統(tǒng)計(jì)分析。 結(jié)果 1電凝大鼠鞏膜淺層靜脈聯(lián)合應(yīng)用絲裂霉素阻斷房水外引流途徑制成模型成功。 2免疫組化法發(fā)現(xiàn)HSP70在正常視網(wǎng)膜細(xì)胞的胞漿和胞核無(wú)陽(yáng)性或極弱陽(yáng)性染色；在實(shí)驗(yàn)組表現(xiàn)在細(xì)胞胞核和／或胞漿的有不同程度的褐色或黃棕色陽(yáng)性染色。HSP70的免疫陽(yáng)性染色主要位于視網(wǎng)膜神經(jīng)節(jié)細(xì)胞層,內(nèi)核層也可見到微弱表達(dá)。 3高眼壓+GGA組與高眼壓組相比在術(shù)后1d時(shí)視網(wǎng)膜內(nèi)HSP70蛋白的表達(dá)略有增加但差異無(wú)統(tǒng)計(jì)學(xué)意義(F=1.106,P=0.352),3d,7d時(shí)視網(wǎng)膜內(nèi)HSP70蛋白的表達(dá)明顯增加,差異有統(tǒng)計(jì)學(xué)意義(F=70.030,P=0.001；F=180.649,P=0.000)；14d,21d組視網(wǎng)膜內(nèi)HSP70蛋白的表達(dá)有所下降但差異仍有統(tǒng)計(jì)學(xué)意義(F=103.086,P=0.001；F=140.811,P=0.000)；28d組視網(wǎng)膜內(nèi)HSP70蛋白的表達(dá)明顯下降與高眼壓組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(F=7.578,P=0.051)。 結(jié)論 1 HSP70在視網(wǎng)膜表達(dá)的部位,主要表達(dá)于節(jié)細(xì)胞層,并少量表達(dá)于內(nèi)核層。 2替普瑞酮能夠有效的促進(jìn)大鼠慢性高眼壓視網(wǎng)膜HSP70的表達(dá)。
[Abstract]:Objective glaucoma is one of the major blinding eye diseases worldwide. Intraocular pressure (IOP) exceeds the bearing range of retinal ganglion cell (rentinal ganglion cells, RGCs), causing RGCs apoptosis to cause progressive irreversible damage to visual function. However, the mechanism of RGCs apoptosis is still unclear. HSP is a special group of proteins produced by biological cells under adverse environmental factors, which can enhance cell viability and protect cells from injury. Heat shock protein (70 (heat shock protein, HSP70) is the most abundant heat shock protein in the central nervous system. Early studies have shown that tipranone (geranglgeranylacetone, GGA) can significantly induce the expression of HSP70 in gastrointestinal mucosa, liver, heart and other tissues. Recent studies have found that a certain dose of tipranone can also induce HSP70 expression in the central nervous system. The external drainage of aqueous humor was blocked by blocking the superficial scleral vein and mitomycin in rats. The intraocular pressure was increased and the pathogenesis of chronic glaucoma was simulated. To observe the effect of tipranone on the expression of HSP70 in the retina of rats with chronic intraocular hypertension, and to provide a new therapeutic approach for the protection of glaucoma optic nerve. Methods 1 the model of chronic high intraocular pressure was established by the combination of superficial scleral vein and mitomycin to block the external drainage of aqueous humor. The intraocular pressure (IOP) was measured by TONO-PEN XL pen type intraocular pressure (IOP) before operation and immediately after operation, 1 day, 3 days, 7 days, 14 days, 21 days and 28 days. 2 the morphological changes of retina were observed by HE staining. 3 the expression of HSP70 protein in normal retina and chronic intraocular pressure (GGA). The expression of HSP70 protein in normal retina, chronic intraocular pressure retina and retina after medication was detected by immunohistochemical method with non-biotin two-step method. The statistical results were expressed as mean 鹵standard deviation. One-Way ANOVA analysis and SPSS13.0 software were used for statistical analysis. Results 1 the model was successfully established by electrocoagulation of superficial scleral vein combined with mitomycin to block the pathway of aqueous humor drainage. 2Immunohistochemical staining showed that HSP70 was not positive or weakly positive in cytoplasm and nucleus of normal retinal cells. In the experimental group, there were different degrees of brown or yellowish brown positive staining in the nucleus and / or cytoplasm of the cells. The immunoreactive staining of HSP70 was mainly located in the retinal ganglion cell layer and weakly expressed in the nuclear layer. 3 the expression of HSP70 protein in the retina of the GGA group was slightly higher than that in the high IOP group on the 1st day after operation, but there was no significant difference (F1. 106 P0. 352). The expression of HSP70 protein in the retina increased significantly at 3 days after 7 days. The difference was statistically significant (FG 70.030 P < 0.001). The expression of HSP70 protein in the retina of 14 d to 21 d group was decreased, but the difference was still significant (F103.086%, F140.811 P0. 000), and the expression of HSP70 protein in the retina of the 14 d group was significantly lower than that of the control group (P < 0. 05%), and the expression of HSP70 protein in the retina was significantly lower than that in the control group. There was no significant difference in the expression of HSP70 protein between the 28 d group and the high IOP group. Conclusion 1 HSP70 is mainly expressed in the ganglion cell layer and a little in the inner layer of the retina. 2 tipranone can effectively promote the expression of retinal HSP70 in rats with chronic intraocular hypertension.
【學(xué)位授予單位】：中國(guó)醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R775

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相關(guān)期刊論文 前1條

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