缺氧诱导因子1α特异性小干扰RNA对早期糖尿病大鼠视网膜白细胞黏附及髓样细胞活性的影响  

The mediation of HIF-α siRNA to the leukocyte adhesion and myeloid cell＇s activity in rat＇s retina under early stage of diabetic retinopathy

作　　者：宋虎平[1] 朱琦[2] 吴琼[1] 宋赛乐 孙红芬[1] 周晓樑 刘晖[1] 

机构地区：[1]西安市第四医院眼科,710004 [2]榆林市第一医院眼科

出　　处：《中华眼科杂志》2015年第5期351-355,共5页Chinese Journal of Ophthalmology

基　　金：陕西省自然科学基金（2011JM4048）;西安市科技发展计划项卧SF1315（2）

摘　　要：目的观察缺氧诱导因子1α特异性小干扰RNA（HIF-1α siRNA）对早期糖尿病大鼠视网膜白细胞黏附及髓样细胞活性的影响。方法实验研究。构建HIF-1α siRNA表达载体pSUPERHl-siHIFlα。通过链脲佐菌素腹腔内注射诱导大鼠糖尿病模型。建模成功后2个月，取27只大鼠随机分为糖尿病＋磷酸盐缓冲液（PBS）玻璃体腔注射组（B组）、糖尿病＋pSUPERHl-siHIFlα玻璃体腔注射转染组（c组）、糖尿病＋pSUPER空载体玻璃体腔注射组（D组），每组9只大鼠。取9只月龄匹配的正常大鼠为对照组（A组）。分别以丫啶橙视网膜白细胞造影和髓过氧化物酶（MPO）酶联免疫吸附试剂盒检测视网膜白细胞黏附数量及髓样细胞活性。四组间均数的差异比较采用单因素方差分析，均数间差异的多重比较采用LSD—t检验。结果丫啶橙视网膜造影检测结果显示，A、B、C、D组视网膜白细胞黏附数量分别为（47．00±3．60）、（155．33±9．01）、（76．00±9．05）、（142．66±10．26）个，差异有统计学意义（F=116．25，P=O．00），C组视网膜白细胞黏附数量显著低于B组（LSD—t检验，P=0．00，95％可信区间为3．56—95．10）。4个组大鼠视网膜上清液中MPO的浓度分别为（17．24±1．13）、（31．32±2．53）、（21．35±1．06）、（31．33±1．26）μg／L，差异有统计学意义（F=58．68，P=0．00），C组视网膜中MPO水平显著低于B组（LSD—t检验，P=0．00，95％可信区间为6．92—13．01）。结论HIF-10lsiRNA对早期糖尿病大鼠视网膜白细胞黏附及髓样细胞活性有一定的调节作用。Objective To investigate the mediation of HIF-α siRNA to the leukocyte adhesion and myeloid cell＇s activity in rat＇s retina at early stage of diabetic retinopathy. Methods Experimental study. HIF-α specific siRNA expression vector pSUPERHI-siHIFlα was constructed by gene recombination. The rat diabetic model was induced by intraperitoneal injection of streptozotocin. After 2 months of diabetes induction, 27 diabetic rats were randomly chosen and assigned to 3 groups, including diabetes and phosphate buffered saline （PBS） vitreous injection group （group B） , diabetes and pSUPERHI-siHIFlα vitreous transfect group （group C） and diabetes and pSUPER-retro vitreous transfect group （group D）. Each group had 9 rats. Nine age matched health rats were chosen as control group （group A）. Retinal leukostasis was quantified with acridine orange leukocyte fluorography. Retinal myeloid cell activity was measured by enzyme linked immunosorbent assay of myeloperoxidase（MPO）. The differences of the mean values among the four groups were analyzed by one-factor analysis of variance. The multiple comparisons of the mean values among the four groups were analyzed by LSD-t analysis. Results According to the results of the acridine orange leukocyte fluorography, the numbers of leukocyte adhesion inthe four groups were （47.00 ± 3.60） , （ 155.33 ± 9.01 ） , （76.00 ±9.05 ） , （ 142.66± 10.26） , respectively. The differences among them were significant （F=116.25, P=0.00）. The number of leukocyte adhesion in the group C was significantly lower than that in group B（LSD-t test, P=0.00,95%CI： 3.56±95.10）.The levels of retinal MPO in the four groups were （ 17.24± 1.13）, （31.32±2.53）, （21.35±1.06）, （31.33± 1.26） μg/L, respectively. The differences among them were significant（F=58.68, P=0.00）. The level of retinal MPO in the group C was significantly lower than that in group B（LSD-t test, P=0.00, 95% CI： 6.92--13.01）. Coneluslons HIF-α siRNA may play a role

关 键 词：糖尿病视网膜病变 细胞黏附 白细胞 髓系细胞 缺氧诱导因子1Α 亚基 RNA 小分子干扰 糖尿病 实验性 

分 类 号：R587.2[医药卫生—内分泌] R774.1[医药卫生—内科学]