摘要目的 观察外源性轴突导向因子-1(netrin-1)对糖尿病(DM)大鼠视网膜Müller细胞活化的影响.方法 健康清洁级雄性Sprague-Dawley大鼠50只,采用随机数字表法随机分为正常对照组(A组)、正常+平衡盐溶液(BSS)组(B组)、正常+netrin-1100μg/ml组(C组)、DM+BSS组(D组)、DM+netrin-1100μg/ml组(E组),每组10只.D、E组大鼠按60 mg/kg的剂量,左下腹腔注射链脲佐菌素诱导DM动物模型.采用免疫组织化学染色法检测大鼠视网膜Müller细胞标志物胶质原纤维酸性蛋白(GFAP)的阳性表达;荧光定量聚合酶链反应检测大鼠视网膜GFAP mRNA的表达.结果 免疫组织化学染色结果显示,A~C组大鼠视网膜仅在神经纤维层及神经节细胞层可见少量GFAP阳性表达.D组大鼠视网膜GFAP阳性表达增多.E组大鼠视网膜GFAP阳性表达较D组减少.A~E组大鼠视网膜GFAP阳性表达、mRNA表达比较,差异有统计学意义(F=203.43、72.91,P=0.00、0.00).与A组比较,D组大鼠视网膜GFAP阳性表达、mRNA表达明显增多,差异有统计学意义(t=-26.01、22.26,P=0.00、0.00).E组大鼠视网膜GFAP阳性表达、mRNA表达较D组明显下降(t=-10.78、3.93,P=0.00、0.00),但仍高于A组(t=7.00、-9.82,P=0.00、0.00),差异均有统计学意义.A、C组大鼠视网膜GFAP阳性表达、mRNA表达比较,差异均无统计学意义(t=-0.29、0.50,P=0.77、0.62).结论 Netrin-1可降低DM大鼠视网膜Müller细胞活化,减少视网膜GFAP表达.

abstractsObjective To observe the effect of netrin-1 on retinal Müller cells in diabetes mellitus (DM) rats. Methods Fifty Sprague-Dawley rats were randomly divided into the normal control group (group A), normal + balanced salt solution (BSS) group (group B), normal+netrin-1 group (group C), DM+BSS group (group D) and DM+netrin-1 group (group E), with 10 rats in each group. DM rats were induced by intraperitoneal injection of Streptozotocin (60 mg/kg). The expression level of glial fibrillary acidic protein (GFAP) on retinal Müller cells was determined by immunohistochemistry, the level of GFAP mRNA was analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction. Results Immunohistochemistry showed that GFAP was distributed in retinal ganglion cells and retinal nerve fiber layer in group A, B and C. Compared to group B, GFAP staining was brighter in the group D. There were significant differences in the expression of GFAP protein and mRNA among groups A-E (F=203.43, 72.91; P=0.00, 0.00), they were higher in group D than group A (t=-26.01, 22.26; P=0.00, 0.00), and group E (t=-10.78, 3.93; P=0.00, 0.00). They were higher in group E than group A (t=7.00, -9.82; P=0.00,0.00). There were no significant differences in between group A and group C (t=-0.29, 0.50; P=0.77, 0.62). Conclusion The expression of GFAP in Müller cells of DM rats could be decreased by injecting netrin-1 into vitreous.