本文選題：糖尿病視網(wǎng)膜病變 + 丙酮酸鈉　； 參考：《復旦大學》2010年碩士論文

【摘要】： 糖尿病視網(wǎng)膜病變(Diabetic Retinopathy, DR)是糖尿病常見微血管并發(fā)癥。隨著糖尿病患者逐漸增多,發(fā)病人群逐漸年輕化,DR成為威脅人群視力的重要疾病。但DR的發(fā)生機制尚未完全明確,缺乏有效的早期治療手段。近年丙酮酸(Pyruvic Acid)作為體內(nèi)糖代謝的樞紐物質(zhì),因其所具有的獨特抗氧化損傷作用而被重視。既往研究證明,丙酮酸鈉(Pyruvate Sodium, NaPyr)在治療糖尿病相關(guān)白內(nèi)障、缺血性心肌細胞損傷、缺血性中樞神經(jīng)系統(tǒng)損傷(Central Nervous System, CNS)等與氧化損傷密切相關(guān)的疾病中已經(jīng)取得較好的療效。因此可推測NaPyr對DR亦具有良好的保護作用。本課題以鏈尿佐菌素(Streptozotocin, STZ)誘導的大鼠糖尿病模型為研究對象,從生物化學、免疫熒光及細胞超微結(jié)構(gòu)觀察等方面來評價NaPyr對DR早期大鼠Muller細胞的保護效果。 目的在體研究NaPyr對STZ大鼠DR的保護作用。 方法用75mg／kg的STZ一次性腹腔注射建立STZ大鼠糖尿病模型,隨機分為糖尿病對照組、口服組和腹腔注射治療組�？诜M于飲水中加入2%NaPyr,腹腔注射治療組隔天向腹腔注射250mg／kg劑量NaPyr滅菌平衡溶液1次,糖尿病對照組隔天向腹腔注射相同體積滅菌生理鹽水1次。分別于1個月、4個月、6個月處死后取視網(wǎng)膜以硫代巴比妥酸法(Thiobarbituric Acid Test, TBA)檢測視網(wǎng)膜組織丙二醛(Malonic Dialdehyde, MDA)的含量,以黃嘌呤氧化酶法(Xanthine Oxidase Test, XOD Test)測定超氧化物歧化酶(Superoxide Dismutase, SOD)含量,免疫熒光法檢測谷氨酰胺酶(GS)、膠質(zhì)纖維酸性蛋白(GFAP)在視網(wǎng)膜上的表達情況及透射電鏡觀察視網(wǎng)膜的超微結(jié)構(gòu)。 結(jié)果在氧化應(yīng)激相關(guān)指標方面,成模1個月時,口服組和對照組MDA均上升,注射給藥組MDA有所下降,具有極顯著差異(P0.001)。成模4個月和6個月時,各組MDA均保持穩(wěn)定,不具有顯著差異(P0.05)。在成模1個月時,各組SOD不具有顯著差異(P0.05)。而4個月時,口服組和注射組SOD比對照組有所升高,具有顯著差異(P0.05)。6個月時,口服組和注射組SOD與對照組相比,不具有顯著差異(P0.05)。在視網(wǎng)膜Muller細胞方面,無論是成模1個月或4個月時,口服與注射組與對照組相比均未發(fā)現(xiàn)GS的染色陽性區(qū)域明顯改變,而口服與注射組GFAP在1個月及4個月時期的染色陽性區(qū)域都較對照組小。成模1個月時,對照組的Muller細胞與神經(jīng)節(jié)細胞超微結(jié)構(gòu)有輕度改變,口服與注射組好于對照組。而成模4個月時,各組神經(jīng)節(jié)細胞及Muller細胞超微結(jié)構(gòu)均受到較重破壞,而口服組受到的破壞輕于對照組及注射組。 結(jié)論NaPyr對DR早期的保護作用可能存在,晚期NaPyr的保護作用尚不確定,有待進一步研究。
[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.
【學位授予單位】：復旦大學
【學位級別】：碩士
【學位授予年份】：2010
【分類號】：R587.2;R774.1

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