替普瑞酮对大鼠慢性高眼压视网膜HSP70表达的影响
发布时间：2018-12-31 18:23
【摘要】： 目的 青光眼是当今世界范围内的主要致盲性眼病之一,由于眼压超过视网膜节细胞(rentinal ganglion cells, RGCs)的承受范围,引起RGCs凋亡造成视功能的进行性不可逆性损害,但RGCs凋亡的机制尚不清楚。HSP是生物体细胞在不良环境因素作用下所产生的一组特殊的蛋白质,能增强细胞存活能力,保护细胞不受或少受伤害。其中热休克蛋白70(heat shock protein, HSP70)是在中枢神经系统含量最丰富的热休克蛋白。早期研究证实抗溃疡药物替普瑞酮(geranglgeranylacetone, GGA)可以明显诱导胃肠粘膜、肝脏、心脏等组织的HSP70表达。新近研究发现一定剂量的替普瑞酮在中枢神经系统也可以诱导HSP70表达。我们通过阻断大鼠巩膜浅层静脉联合丝裂霉素的方法阻断房水的外引流途径,使其眼压升高,模拟慢性青光眼的发病过程。观察替普瑞酮对大鼠慢性高眼压视网膜HSP70表达的影响,为青光眼视神经保护提供新的治疗思路。 方法 1慢性高眼压模型的制备 热凝大鼠巩膜浅层静脉联合应用丝裂霉素阻断房水外引流途径制成模型；TONO-PEN XL型笔式眼压计测量术前,术后即刻,1d,3d,7d,14d,21d,28d眼压。 2 HE染色观察视网膜形态学变化。 3正常视网膜,慢性高眼压视网膜,慢性高眼压+GGA后视网膜HSP70蛋白表达水平的初步研究。 应用非生物素二步法免疫组化法检测正常视网膜,慢性高眼压视网膜,用药后视网膜中HSP70蛋白表达的部位和程度。统计结果均以均数±标准差表示,组间比较应用One-Way ANOVA分析,均用SPSS13.0软件进行统计分析。 结果 1电凝大鼠巩膜浅层静脉联合应用丝裂霉素阻断房水外引流途径制成模型成功。 2免疫组化法发现HSP70在正常视网膜细胞的胞浆和胞核无阳性或极弱阳性染色；在实验组表现在细胞胞核和／或胞浆的有不同程度的褐色或黄棕色阳性染色。HSP70的免疫阳性染色主要位于视网膜神经节细胞层,内核层也可见到微弱表达。 3高眼压+GGA组与高眼压组相比在术后1d时视网膜内HSP70蛋白的表达略有增加但差异无统计学意义(F=1.106,P=0.352),3d,7d时视网膜内HSP70蛋白的表达明显增加,差异有统计学意义(F=70.030,P=0.001；F=180.649,P=0.000)；14d,21d组视网膜内HSP70蛋白的表达有所下降但差异仍有统计学意义(F=103.086,P=0.001；F=140.811,P=0.000)；28d组视网膜内HSP70蛋白的表达明显下降与高眼压组比较差异无统计学意义(F=7.578,P=0.051)。 结论 1 HSP70在视网膜表达的部位,主要表达于节细胞层,并少量表达于内核层。 2替普瑞酮能够有效的促进大鼠慢性高眼压视网膜HSP70的表达。
[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 卤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.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R775
本文编号：2396947
[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 卤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.
【学位授予单位】：中国医科大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R775
【参考文献】
相关期刊论文 前1条
1 肖红,王泉云,廖刃,张兰,梁茂植;氯胺酮-咪唑安定对感染性休克大鼠血清肿瘤坏死因子-α及心肌热休克蛋白70表达的影响[J];临床麻醉学杂志;2001年09期
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