吡哆胺对糖尿病大鼠视网膜AGEs及其受体基因表达的影响

发布时间：2018-11-20 12:46

【摘要】：目的：研究吡哆胺对糖尿病大鼠视网膜中晚期糖基化终末产物(AGEs)及其受体(RAGE)表达的影响，探讨吡哆胺在保护糖尿病大鼠视网膜中的作用。方法：将6周龄（清洁级）雄性SD大鼠以单次腹腔注射链脲佐菌素(STZ)诱发为1型糖尿病。实验大鼠分为4组，①吡哆胺处理组（PM组）：STZ注射72h后采用吡哆胺（200mg·kg-1·d-1）灌胃；②氨基胍处理组（AG组）：STZ注射72h后采用氨基胍(100mg·kg-1·d-1)灌胃；③对照组（DM组）：STZ注射72h后采用蒸馏水(2ml/d)灌胃；④正常对照组（NC组）：正常大鼠柠檬酸缓冲液注射72h后采用蒸馏水(2ml/d)灌胃。治疗4周和12周后取大鼠血清和视网膜，采用ELISA法测定AGEs含量，使用免疫荧光染色和实时荧光定量PCR观察视网膜中RAGE的分布和表达情况。结果：治疗4周时，ELISA检测各组血清和视网膜AGEs含量结果显示：PM组为2.430±0.096ug/l，2.406±0.118ug/l；AG组为3.360±0.110ug/l，3.324±0.110ug/l；DM组为3.789±0.128ug/l，3.811±0.130ug/l；PM组与AG组和DM组比较差异有统计学意义（P0.01）。治疗12周时，各组血清和视网膜AGEs含量结果： PM组为2.934±0.061ug/l，2.909±0.055ug/l；AG组为3.888±0.080ug/l，3.812±0.120ug/l；DM组为4.327±0.100ug/l，4.367±0.103ug/l；PM组与AG组和DM组比较差异有统计学意义（P0.01）。12周时各组AGEs含量比4周时明显增加（P0.01）。免疫荧光染色检测发现RAGE表达于视网膜色素上皮层至神经节细胞层，治疗4周时OD值分别为PM组1.72±0.04，AG组1.56±0.04，DM组1.30±0.02，，PM组与AG组和DM组比较差异有统计学意义（P0.01）。治疗12周时OD值分别为PM组1.52±0.22，AG组1.37±0.26，DM组1.00±0.02，PM组与AG组和DM组比较差异有统计学意义（P0.01）。12周时各组RAGE表达量明显高于4周时（P0.01）。实时荧光定量PCR对RAGE mRNA的检测结果趋势同免疫荧光染色结果一致。结论：使用吡哆胺治疗能降低糖尿病大鼠血清和视网膜AGEs的含量，并下调其受体RAGE的表达，减轻二者对视网膜产生的毒性损害，具有保护视网膜的作用。
[Abstract]:Aim: to study the effect of pyridoxamine on the expression of (AGEs) and its receptor (RAGE) in the late stage of diabetic rat retina, and to explore the role of pyridoxamine in protecting the retina of diabetic rats. Methods: 6 weeks old male SD rats were induced into type 1 diabetes mellitus by single intraperitoneal injection of streptozotocin (STZ). The experimental rats were divided into 4 groups. 1 the rats in the PM group were treated with pyridoxamine (200mg kg-1 d-1) for 72 h after): STZ injection. 2Aminoguanidine treated group (AG group) was treated with 100mg kg-1 d-1 after 72 h of): STZ injection, and control group (DM group) was treated with distilled water (2ml/d) 72 h after): STZ injection. 4 normal control group (NC group): normal rats were treated with distilled water (2ml/d) for 72 h after injection of citric acid buffer. After 4 and 12 weeks of treatment, the serum and retina of the rats were taken. The content of AGEs was measured by ELISA method. The distribution and expression of RAGE in the retina were observed by immunofluorescence staining and real-time fluorescence quantitative PCR. Results: at 4 weeks after treatment, the serum and retinal AGEs levels in the PM group were 2.430 卤0.096ugP / L 2.406 卤0.118ugP / L + AG = 3.360 卤0.110ugP / L = 3.324 卤0.110ugP / L, respectively. There was significant difference between DM group and AG group and DM group (3.789 卤0.128ugP / L = 3.811 卤0.130ugP / L) (P0.01). After 12 weeks of treatment, the serum and retinal AGEs levels in the PM group were 2.934 卤0.061ugP / L 2.909 卤0.055ugP / L AG, 3.888 卤0.080ugP / L = 3.812 卤0.120ugP / L, and 4.327 卤0.100ugP / L = 4.367 卤0.103ugP / l, respectively, in the PM group (4.327 卤0.100ugP / L = 4.367 卤0.103ugP / l). The content of AGEs in PM group was significantly higher than that in AG group and DM group at 12 weeks (P0.01). Immunofluorescence staining showed that RAGE was expressed in the retinal pigment epithelium layer to the ganglion cell layer. The OD values at 4 weeks after treatment were 1.72 卤0.04 AG in PM group 1.56 卤0.04 渭 m in DM group and 1.30 卤0.02 in DM group, respectively. There was significant difference between PM group and AG group and DM group (P0.01). At 12 weeks after treatment, the OD values of PM group were 1.52 卤0.22 AG group 1.37 卤0.26 渭 g DM group 1.00 卤0.02, respectively. The expression of RAGE in PM group was significantly higher than that in AG group and DM group at 12 weeks (P0.01). The trend of RAGE mRNA detection by real-time fluorescence quantitative PCR was consistent with that of immunofluorescence staining. Conclusion: the treatment of pyridoxamine can reduce the content of AGEs in serum and retina of diabetic rats, and down-regulate the expression of RAGE receptor, alleviate the toxic damage to retina and protect the retina.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R774.1

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