葛根素对糖尿病大鼠的视网膜中糖基化修饰蛋白的影响

Effect of puerarin on glycosylated modified proteins in retina of diabetic rats

目的研究葛根素对糖尿病大鼠视网膜中AGEs-修饰蛋白的影响。方法以2%链脲佐菌素55 mg·kg^(-1)腹腔注射建立糖尿病大鼠模型。按照体重将大鼠随机分为6组:正常^(-1)组与正常-2组(1代表1个月,2代表3个月),糖尿病^(-1)组与模型-2组,实验^(-1)组与实验-2组(葛根素分别干预1,3个月),每组25只。大鼠造模成功后,实验组予以葛根素80 mg·kg^(-1)·d^(-1)灌胃,模型组和正常组大鼠给予0.9%NaCl灌胃。以酶联免疫吸附法测定视网膜中AGEs-修饰蛋白的总浓度;以免疫印迹法测定视网膜中AGEs-修饰蛋白表达水平。结果正常-2组、模型-2组、实验-2组的视网膜AGEs-修饰蛋白的总质量浓度分别是(78.14±3.80),(338.33±25.31),(269.42±20.58)mg·L^(-1);实验-2组和模型-2组与正常组比较,差异均有统计学意义(均P<0.05);实验-2组与模型-2组比较,差异有统计学意义(P<0.05)。模型-2组和实验-2组与正常组比较,4种差异性表达的AGEs-修饰蛋白的表达水平上调;而实验-2组与模型-2组比较,其表达水平下调,差异均有统计学意义(均P<0.05)。结论葛根素干预可以改善糖尿病大鼠视网膜的病理损害,其机制可能与葛根素抑制AGEs-修饰蛋白的生成、减少AGEs-修饰蛋白在视网膜组织中蓄积有关,但其强度与作用时间相关。

Objective To study the effect of puerarin on the expression of advanced glycated end-products（ AGEs）-modified proteins in retina of diabetic rats. Methods A diabetic rat model was established by intraperitoneal injection of 2% streptozotocin 55 mg ·kg^-1. The rats were randomly divided into 6 groups,25 rats in each group： normal^-1 group,normal-2 group（ administration for 1 month,3 months respectively）;model^-1 group,model-2 group（ diabetic 1 month,3 months）,experimental^-1 group,experimental-2 group（ puerarin 80 mg ·kg^（-1）·d^-1 intervention 1 month,3 months,by oral administration per day）; the rats of normal group and model group were given 0. 9% Na Cl,after setting up the diabetic rat model. The total concentrations of AGEs-modified protein in retina were determined by enzyme linked immunosorbent assay. The expression level of AGEs-modified protein in retina was determined by Western blot. Results The total concentrations of AGEs-modified proteins of the normal-2 group,the model-2 group and the experimental-2 group were（ 78. 14 ± 3. 80）,（ 338. 33 ± 25. 31）,（ 269. 42 ± 20. 58）mg·L^-1,respectively; there were statistically significant differences between the normal group and the model group,the experimental group（ all P〈0. 05）; the difference was statistically significant between the model-2 group and the experimental-2 group. The expressional levels of the four differentially expressed AGEs-modified proteins in the model-2 group and the experimental-2 group were up-regulated with statistically significant（ P〈0. 05）; compared with the model-2 group,the expressional levels of the experimental-2 group were down-regulated with statistically significant（ P〈0. 05）. Conclusion Puerarin intervention can improve retinal pathological damage for diabetic rats.The mechanism may be related to puerarin inhibiting the production of AGEs-modified proteins,reducing AGEs-modified proteins accumulation in retinal tissues,and the intensity of effect was closely related to working times.