摘要
目的 探讨葡萄糖转运蛋白GLUT1/4和Sirtuins在糖尿病大鼠视网膜中的表达。方法 取8周龄健康雄性SD大鼠20只随机分为正常对照组和糖尿病组,糖尿病组大鼠按照60 mg·kg^(-1)的剂量采用一次性腹腔注射链脲佐菌素诱导糖尿病模型,正常对照组大鼠注射等量的溶剂。每隔2周检测大鼠体重和血糖。造模后12周,应用彩色多普勒超声检测大鼠视网膜中央动脉(CRA)血流参数;使用戊巴比妥钠麻醉大鼠后摘取眼球,采用HE染色观察大鼠视网膜组织的病理变化,免疫组织化学染色、Western blot和qRT-PCR分别检测大鼠视网膜中GLUT1/4和Sirtuins mRNA的表达情况。结果 造模后12周,与正常对照组相比,糖尿病组大鼠CRA的收缩期峰值流速和舒张末期流速均显著降低(均为P<0.001);阻力指数和搏动指数无明显改变(均为P>0.05)。造模后12周HE染色结果显示,正常对照组大鼠视网膜组织各层结构清晰,细胞排列紧密、规则,未发现明显的病理改变;糖尿病组大鼠视网膜组织厚度变薄,各层界限模糊,结构紊乱,细胞数量减少。造模后12周免疫组织化学染色结果显示,在大鼠视网膜中,GLUT1主要位于视网膜色素上皮层,GLUT4位于神经节细胞层、内丛状层及光感受器层;Western blot检测结果显示,糖尿病组大鼠视网膜中GLUT1及GLUT4蛋白相对表达量均低于正常对照组(均为P<0.05),糖尿病组大鼠视网膜中SIRT1-SIRT7蛋白相对表达量均低于正常对照组(均为P<0.05)。qRT-PCR检测结果显示,与正常对照组相比,糖尿病组大鼠视网膜中的SIRT1-SIRT7 mRNA相对表达量均下降,差异均有统计学意义(均为P<0.01)。结论 糖尿病可引起大鼠视网膜组织中GLUT1/4和Sirtuins的表达发生改变,GLUT1/4和Sirtuins可能参与了DR的发生与发展。
Objective To explore the changes in the expression of glucose transporters 1/4(GLUT1/4)and Sirtuins in the retina of rats with diabetes.Methods Twenty 8-week-old healthy male Sprague-Dawley rats were randomly divided into normal control and diabetic groups.Rats in the diabetic group received a disposable intraperitoneal injection of 60 mg·kg^(-1) streptozotocin to induce the diabetes model,while rats in the normal control group were injected with an equivalent amount of solvent.Body weight and blood glucose were measured at 2-week intervals.At 12 weeks after modeling,color Doppler ultrasound was applied to detect blood flow parameters in the central retinal artery(CRA)of rats;after anesthetizing rats with sodium pentobarbital,eyeballs were harvested,and the pathological changes of rat retinal tissue were observed by hematoxylin&eosin(HE)staining.The expression of messenger ribonucleic acid(mRNA)for GLUT 1/4 and Sirtuins in the retina of rats were detected by immunohistochemical staining,Western blot and quantitative of reverse transcription polymerase chain reaction(qRT-PCR),respectively.Results At 12 weeks after modeling,compared with the normal control group,peak systolic velocity and end diastolic velocity were significantly lower in CRA of rats in the diabetic group(both P<0.001);there were no significant differences in resistance index and pulsatility index(both P>0.05).The HE staining results at 12 weeks after modeling showed that rats in the normal control group had clear structure in each layer of retinal tissues,closely and regularly arranged cells,and no obvious pathological changes;rats in the diabetic group showed decreased retinal thickness,blurred boundary of each layer,disordered structure and reduced cell number.Immunohistochemical staining at 12 weeks after modeling showed that GLUT 1 was mainly located in the retinal pigment epithelial layer of rats,and GLUT 4 was located in the ganglion cell layer,inner plexiform layer and photoreceptor layer.Western blot results showed that the relative expre
作者
白文帆
郭玉
伏等弟
罗明秀
芦晓红
姚青
BAI Wenfan;GUO Yu;FU Dengdi;LUO Mingxiu;LU Xiaohong;YAO Qing(Department of Biochemistry and Molecular Biology,School of Basic Medicine,Ningxia Medical University,Yinchuan 750004,Ningxia Autonomous Region,China)
出处
《眼科新进展》
CAS
北大核心
2024年第4期270-274,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金项目(编号:82060819)
宁夏回族自治区自然科学基金项目(编号:2020AAC02024)。
