糖康对糖尿病视网膜病变的影响及其作用机制研究
发布时间：2019-03-22 10:35
【摘要】： 目的:从视网膜微血管形态学和细胞分子两个水平,观察糖康对糖尿病大鼠视网膜病变的影响,并探讨其具体作用机制,为糖康防治糖尿病视网膜病变提供理论依据。方法:采用腹腔注射链脲佐菌素复制糖尿病模型,并随机分为正常对照组、模型组、糖康高、中和低剂量组和达纳康组,每日灌胃,定期检测大鼠体重和非空腹血糖。第12周末,消化铺片法观察视网膜微血管形态学改变;化学比色法检测各组大鼠血清及视网膜组织中超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。Western blot检测NF-κB亚基P65在视网膜组织细胞核内的含量。半定量RT-PCR检测其上、下游靶基因IL-1β、Bax和VEGF的表达水平。结果:经STZ诱发的糖尿病大鼠模型,造模一周左右即出现DM典型的“三多”症状。糖康高、中剂量组和与模型组相比,上述症状轻于模型组,而糖康低剂量组和达纳康组与同期模型组相比无明显变化;模型组与各治疗组大鼠较同期正常组大鼠体重明显降低(P0.01);正常组大鼠血糖保持稳定,模型组与各治疗组大鼠较同期正常组大鼠组血糖明显升高(P0.01);与正常组比较,模型组大鼠视网膜呈现较多的无细胞毛细血管条索数(约为正常组的12.16倍)。与模型组比较,糖康高、中、低剂量组和达纳康组无细胞毛细血管条索数明显减少(P0.01);与模型组比较,各治疗组大鼠血清及视网膜组织的SOD活性均不同程度升高,MDA含量均不同程度降低,以糖康高剂量组效果最为显著(P0.01);与正常对照组比较,模型组NF-κBP65蛋白表达水平明显上调(P0.01),各治疗组NF-κBP65蛋白在视网膜组织的表达不同程度降低,以糖康高剂量组最为显著;与正常组比较,模型组大鼠视网膜IL-1β、VEGF和Bax的mRNA表达明显上调(P0.01)。与模型组比较,各治疗组大鼠视网膜IL-1β、VEGF和Bax的mRNA表达均不同程度降低,以糖康高剂量组最为显著。结论:用STZ诱发糖尿病大鼠模型是一种快速、简便、有效的动物模型制备方法,STZ最佳用药浓度为50 mg/kg;糖康能够改善DM大鼠的诸多糖尿病症状,但对血糖和体重改善不明显;糖康能够明显抑制糖尿病诱发的视网膜微血管细胞的凋亡和无细胞毛细血管条索数的增加,对糖尿病大鼠视网膜微血管具有一定的保护作用;糖康能够显著增加糖尿病大鼠血清及视网膜SOD活性,降低MDA含量,说明该药物具有清除糖尿病大鼠体内自由基,增强机体抗氧化防御能力,调节和改善自由基代谢的作用;糖康对糖尿病大鼠视网膜的保护作用可能与减少NF-κBP65在细胞核内的蛋白表达有关;糖康对糖尿病大鼠视网膜的保护作用可能与下调IL-1β、VEGF和Bax的基因表达有关。
[Abstract]:Objective: To observe the effect of Tangkang on the diabetic retinopathy in diabetic rats from two levels of retinal microvessel morphology and cell molecule, and to provide a theoretical basis for the prevention and treatment of diabetic retinopathy. Methods: The model of diabetes mellitus was duplicated by intraperitoneal injection of streptozoin. The rats were randomly divided into the normal control group, the model group, the sugar-kang group, the middle and low-dose group and the Dana-kang group, and the body weight and the non-fasting blood sugar were detected on a regular basis. The changes of the microvessel morphology of the retina were observed at the end of the 12th week, and the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in the serum and retinal tissues of each group were detected by a chemical colorimetric method. Western blot was used to detect the content of NF-VIB subunit P65 in the nucleus of the retinal tissue. The expression levels of IL-1, Bax and VEGF were detected by semi-quantitative RT-PCR. Results: The model of diabetic rats induced by STZ was the typical "Sando" of DM in about one week. The above-mentioned symptoms were mild in the model group compared with the model group, while the low-dose group and the Dana-kang group did not change significantly compared with the model group, and the weight of rats in the model group and the treatment group was significantly lower than that in the normal group (P0.01). In the normal group, the blood glucose remained stable, and the blood sugar in the model group and the control group was significantly higher than that in the normal group (P 0.01). Compared with the normal group, the rat retina of the model group exhibited more cell-free capillary strips (about 12.16 times the normal group). Compared with the model group, there was a significant decrease in the number of cell-free capillary cells in the high, middle and low-dose group and in the Dana-kang group (P0.01). Compared with the model group, the activity of SOD in the serum and retinal tissues of the rats in the treatment group increased significantly, and the content of MDA decreased. Compared with the normal control group, the expression of NF-BBP65 protein in the model group was up-regulated (P0.01), and the expression of NF-BBP65 protein in each treatment group was lower in the retinal tissue, and the group was the most significant in the high-dose group. Compared with the normal group, the expression of IL-1, VEGF and Bax in rat retina was up-regulated (P0.01). Compared with the model group, the expression of IL-1, VEGF and Bax in the retina of each treatment group was decreased to a different extent, and the group was the most significant in the high-dose group. Conclusion: The model of STZ-induced diabetic rats is a fast, simple and effective method for preparing animal model. The optimal concentration of STZ is 50 mg/ kg. The sugar-kang can obviously inhibit the apoptosis of the retinal microvessel cells induced by diabetes and the increase of the number of the cell-free capillary strips, have a certain protective effect on the diabetic rat retina microvessel, and can remarkably increase the SOD activity of the serum and the retina of the diabetic rats, The content of MDA was decreased, which indicated that the drug had the effects of scavenging free radicals in diabetic rats, enhancing the anti-oxidative defense ability of the body, regulating and improving the free-radical metabolism, and the protective effect of the sugar-kang on the retina of the diabetic rats may be related to the reduction of the expression of the NF-BBP65 in the nucleus of the nucleus; The protective effect of Tangkang on the retina of diabetic rats may be related to the down-regulation of the expression of IL-1, VEGF and Bax.
【学位授予单位】：新疆农业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R587.1;R774.1
本文编号：2445517
[Abstract]:Objective: To observe the effect of Tangkang on the diabetic retinopathy in diabetic rats from two levels of retinal microvessel morphology and cell molecule, and to provide a theoretical basis for the prevention and treatment of diabetic retinopathy. Methods: The model of diabetes mellitus was duplicated by intraperitoneal injection of streptozoin. The rats were randomly divided into the normal control group, the model group, the sugar-kang group, the middle and low-dose group and the Dana-kang group, and the body weight and the non-fasting blood sugar were detected on a regular basis. The changes of the microvessel morphology of the retina were observed at the end of the 12th week, and the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in the serum and retinal tissues of each group were detected by a chemical colorimetric method. Western blot was used to detect the content of NF-VIB subunit P65 in the nucleus of the retinal tissue. The expression levels of IL-1, Bax and VEGF were detected by semi-quantitative RT-PCR. Results: The model of diabetic rats induced by STZ was the typical "Sando" of DM in about one week. The above-mentioned symptoms were mild in the model group compared with the model group, while the low-dose group and the Dana-kang group did not change significantly compared with the model group, and the weight of rats in the model group and the treatment group was significantly lower than that in the normal group (P0.01). In the normal group, the blood glucose remained stable, and the blood sugar in the model group and the control group was significantly higher than that in the normal group (P 0.01). Compared with the normal group, the rat retina of the model group exhibited more cell-free capillary strips (about 12.16 times the normal group). Compared with the model group, there was a significant decrease in the number of cell-free capillary cells in the high, middle and low-dose group and in the Dana-kang group (P0.01). Compared with the model group, the activity of SOD in the serum and retinal tissues of the rats in the treatment group increased significantly, and the content of MDA decreased. Compared with the normal control group, the expression of NF-BBP65 protein in the model group was up-regulated (P0.01), and the expression of NF-BBP65 protein in each treatment group was lower in the retinal tissue, and the group was the most significant in the high-dose group. Compared with the normal group, the expression of IL-1, VEGF and Bax in rat retina was up-regulated (P0.01). Compared with the model group, the expression of IL-1, VEGF and Bax in the retina of each treatment group was decreased to a different extent, and the group was the most significant in the high-dose group. Conclusion: The model of STZ-induced diabetic rats is a fast, simple and effective method for preparing animal model. The optimal concentration of STZ is 50 mg/ kg. The sugar-kang can obviously inhibit the apoptosis of the retinal microvessel cells induced by diabetes and the increase of the number of the cell-free capillary strips, have a certain protective effect on the diabetic rat retina microvessel, and can remarkably increase the SOD activity of the serum and the retina of the diabetic rats, The content of MDA was decreased, which indicated that the drug had the effects of scavenging free radicals in diabetic rats, enhancing the anti-oxidative defense ability of the body, regulating and improving the free-radical metabolism, and the protective effect of the sugar-kang on the retina of the diabetic rats may be related to the reduction of the expression of the NF-BBP65 in the nucleus of the nucleus; The protective effect of Tangkang on the retina of diabetic rats may be related to the down-regulation of the expression of IL-1, VEGF and Bax.
【学位授予单位】：新疆农业大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R587.1;R774.1
【引证文献】
相关博士学位论文 前1条
1 郭艺娟;益气养血通络法干预糖尿病视网膜病变的临床与实验研究[D];中国中医科学院;2012年
，本文编号：2445517
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