丙酮酸钠对糖尿病大鼠视网膜保护作用的研究

发布时间：2018-05-03 19:47

本文选题：糖尿病视网膜病变 + 丙酮酸钠　；参考：《复旦大学》2010年硕士论文

【摘要】： 糖尿病视网膜病变(Diabetic Retinopathy, DR)是糖尿病常见微血管并发症。随着糖尿病患者逐渐增多,发病人群逐渐年轻化,DR成为威胁人群视力的重要疾病。但DR的发生机制尚未完全明确,缺乏有效的早期治疗手段。近年丙酮酸(Pyruvic Acid)作为体内糖代谢的枢纽物质,因其所具有的独特抗氧化损伤作用而被重视。既往研究证明,丙酮酸钠(Pyruvate Sodium, NaPyr)在治疗糖尿病相关白内障、缺血性心肌细胞损伤、缺血性中枢神经系统损伤(Central Nervous System, CNS)等与氧化损伤密切相关的疾病中已经取得较好的疗效。因此可推测NaPyr对DR亦具有良好的保护作用。本课题以链尿佐菌素(Streptozotocin, STZ)诱导的大鼠糖尿病模型为研究对象,从生物化学、免疫荧光及细胞超微结构观察等方面来评价NaPyr对DR早期大鼠Muller细胞的保护效果。目的在体研究NaPyr对STZ大鼠DR的保护作用。方法用75mg／kg的STZ一次性腹腔注射建立STZ大鼠糖尿病模型,随机分为糖尿病对照组、口服组和腹腔注射治疗组。口服组于饮水中加入2%NaPyr,腹腔注射治疗组隔天向腹腔注射250mg／kg剂量NaPyr灭菌平衡溶液1次,糖尿病对照组隔天向腹腔注射相同体积灭菌生理盐水1次。分别于1个月、4个月、6个月处死后取视网膜以硫代巴比妥酸法(Thiobarbituric Acid Test, TBA)检测视网膜组织丙二醛(Malonic Dialdehyde, MDA)的含量,以黄嘌呤氧化酶法(Xanthine Oxidase Test, XOD Test)测定超氧化物歧化酶(Superoxide Dismutase, SOD)含量,免疫荧光法检测谷氨酰胺酶(GS)、胶质纤维酸性蛋白(GFAP)在视网膜上的表达情况及透射电镜观察视网膜的超微结构。结果在氧化应激相关指标方面,成模1个月时,口服组和对照组MDA均上升,注射给药组MDA有所下降,具有极显著差异(P0.001)。成模4个月和6个月时,各组MDA均保持稳定,不具有显著差异(P0.05)。在成模1个月时,各组SOD不具有显著差异(P0.05)。而4个月时,口服组和注射组SOD比对照组有所升高,具有显著差异(P0.05)。6个月时,口服组和注射组SOD与对照组相比,不具有显著差异(P0.05)。在视网膜Muller细胞方面,无论是成模1个月或4个月时,口服与注射组与对照组相比均未发现GS的染色阳性区域明显改变,而口服与注射组GFAP在1个月及4个月时期的染色阳性区域都较对照组小。成模1个月时,对照组的Muller细胞与神经节细胞超微结构有轻度改变,口服与注射组好于对照组。而成模4个月时,各组神经节细胞及Muller细胞超微结构均受到较重破坏,而口服组受到的破坏轻于对照组及注射组。结论NaPyr对DR早期的保护作用可能存在,晚期NaPyr的保护作用尚不确定,有待进一步研究。
[Abstract]:Diabetic retinopathy (DRR) is a common microvascular complication of diabetes mellitus. With the increasing number of diabetic patients, the onset of diabetes mellitus (Dr) has become an important disease threatening the visual acuity of the population. However, the pathogenesis of Dr is not completely clear, and lack of effective early treatment. Pyruvic acid (Pyruvic Acid), as a pivotal substance of glucose metabolism in vivo, has been paid more attention recently because of its unique antioxidant damage. Previous studies have shown that Pyruvate Sodium (NaPyrn) has been effective in the treatment of diabetes-related cataract, ischemic myocardial cell injury, ischemic central nervous system injury and other diseases closely related to oxidative injury. Therefore, it can be inferred that NaPyr also has a good protective effect on Dr. In this study, streptozotocin (STZ) induced diabetic rat model was used to evaluate the protective effect of NaPyr on Muller cells in early Dr rats from the aspects of biochemistry, immunofluorescence and ultrastructure observation. Objective to study the protective effect of NaPyr on Dr of STZ rats in vivo. Methods Diabetic model of STZ rats was established by intraperitoneal injection of 75mg/kg (STZ). The rats were randomly divided into three groups: diabetic control group, oral group and intraperitoneal injection group. In the oral group, 2 NaPyrs were added to the drinking water. In the treatment group, the 250mg/kg dose of NaPyr sterilizing equilibrium solution was injected intraperitoneally every other day, and the diabetic control group was injected with the same volume of sterilized saline once every other day. After 1 month, 4 months and 6 months of death, the content of malonic malondialdehyde (MDA) in retina was detected by thiobarbituric Acid Test, TBA), and the content of superoxide dismutase (SOD) was determined by xanthine Oxidase Test, XOD Test). The expression of glutamine glutathione (GSN) and glial fibrillary acidic protein (GFAP) on the retina were detected by immunofluorescence and the ultrastructure of the retina was observed by transmission electron microscope (TEM). Results in the index of oxidative stress, MDA increased in both oral group and control group after 1 month of model formation, and MDA decreased in injection group, with a significant difference (P 0.001). After 4 months and 6 months of model formation, the MDA of each group remained stable, and there was no significant difference (P 0.05). At 1 month, there was no significant difference in SOD in each group (P 0.05). At 4 months, the SOD of the oral group and the injection group was higher than that of the control group, and the difference was significant (P 0.05). At 6 months, the SOD of the oral group and the injection group was not significantly different from that of the control group (P 0.05). In retinal Muller cells, no significant changes were found in the positive areas of GS in the oral and injection groups compared with those in the control group after 1 or 4 months of model formation. The positive areas of GFAP in oral and injection groups were smaller than those in control group at 1 and 4 months. The ultrastructure of Muller cells and ganglion cells in the control group was slightly changed after 1 month of model formation, which was better in the oral and injection groups than in the control group. However, the ultrastructure of ganglion cells and Muller cells in each group was severely damaged after 4 months of model formation, but the damage in oral group was less than that in control group and injection group. Conclusion the early protective effect of NaPyr on Dr may exist, but the protective effect of late NaPyr is uncertain, which needs further study.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R587.2;R774.1

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