摘要目的 观察血管内皮生长抑制因子(VEGI)及其相关因子在糖尿病(DM)大鼠外周血、玻璃体液及视网膜组织中的表达,探讨VEGI在糖尿病视网膜病变(DR)发病机制中的作用.方法 70只6周龄雄性Wistar大鼠,随机分为空白对照组(10只),DM 1、3、6个月组(各20只).DM大鼠模型建立后,分别于1、3、6个月时取大鼠外周血、玻璃体液、全眼球.酶联免疫吸附测定法检测肿瘤坏死因子样配体1/血管内皮生长抑制因子251(TL1A/VEGI 251)、血管内皮生长因子(VEGF)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)在血清及玻璃体液浓度,石蜡切片免疫组织化学法检测各因子在视网膜组织中的表达,苏木素-伊红(HE)染色评价各组大鼠DR进程.比较空白对照组和DM实验组间大鼠血清、玻璃体液及视网膜组织中TL1A、VEGF、TNF-α、IL-1β的表达差异.结果 分析采用单因素方差分析、独立样本f检验和最小显著差法检验.结果 空白对照组、DM 1、3、6个月组大鼠血清TL1A浓度分别为(92.09±2.05)、(118.36±8.30)、(85.90±7.51)、(78.90±4.88) ng/L,组间血清TL1A浓度比较,差异有统计学意义(F=77.405,P＜0.05).从空白对照组到DM 1、3、6个月组,大鼠血清TNF-α、IL-1β浓度均呈升高趋势,4组间血清TNF-α、IL-1β浓度比较,差异有统计学意义(F=3.508、15.416,P＜0.05).VEGF浓度在DM 1个月时升高,DM 3个月时下降,DM 6个月时再次升高,但4组间VEGF浓度比较,差异无统计学意义(F=1.242,P＞0.05).各组大鼠玻璃体液TL1A浓度分别为(91.50±8.18)、(67.03±6.74)、(47.44±4.92)、(46.01±4.62) ng/L,组间TL1A浓度比较,差异有统计学意义(F=114.777,P＜0.05).从空白对照组到DM 1、3、6个月组,大鼠玻璃体液VEGF、TNF-α、IL-1β浓度均呈升高趋势,4组间VEGF、TNF-α、IL-1β比较,差异有统计学意义(F=8.816、4.392、3.635,P＜0.05).免疫组织化学检测结果显示,DM 1、3个月组大鼠视网膜TL1A表达吸光度[A,旧称光密度(OD)]值均较空白对照组降低(t=6.851、6.066,P＜0.05),但DM 6个月组与空白对照组的TL1A表达A值比较,差异无统计学意义(t=1.401,P＞0.05);DM各组大鼠视网膜VEGF和TNF-α表达A值均较空白对照组显著升高(tVEGF=-4.709、-16.406、-9.228,P＜0.05;t TNF.=-4.703、-6.583、-17.762,P＜0.05);DM 1、6个月组大鼠视网膜IL-1β表达A值均较空白对照组显著升高(t=-4.108、-3.495,P＜0.05),但DM 3个月组与空白对照组IL-1β表达A值比较,差异无统计学意义(t=-0.997,P＞0.05).HE染色结果显示,空白对照组与DM 1个月组大鼠视网膜组织结构基本正常;DM 3个月组大鼠视网膜组织水肿,细胞排列紊乱;DM 6个月组大鼠视网膜组织明显水肿,神经节细胞核染色加深,内丛状层可见大量新生血管,内核层细胞排列紊乱、水肿,可见大量脂肪空泡,视网膜各层结构不清晰.结论 VEGI可能通过与VEGF、TNF-α、IL-1β等因子的相互作用参与DR的发生发展过程.

abstractsObjective To observe the expression of vascular endothelial growth inhibitor VEGI.TL1A),vascular endothelial growth factor (VEGF),tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) in diabetes rats' serum,vitreous and retina,and discuss the role of VEGI in the pathogenesis of diabetic retinopathy (DR).Methods A total of p70 adult male Wistar rats were randomly divided into 4 groups,the control group (10 rats),the diabetes mellitus (DM) 1 month group (20 rats),the DM 3 month group (20 rats) and the DM 6 month group (20 rats).Cytokines of serum and vitreous were determined by enzyme-linked immunosorbent assay (ELISA),and the concentrations of the cytokines in the retina were determined by immunohistochemistry on paraffin retinal sections.Hematoxylin-eosin (HE) staining of retina was used to estimate the pathological change of DR.The results were analyzed by one-way analysis of variances,independent samples t-test and LSD test.Results The serum TL1A levels of the control group,the DM 1 month group,the DM 3 month group and the DM 6 month group rats were (92.09 ±2.05),(118.364±8.30),(85.904±7.51) and (78.904±4.88) ng/L respectively,the level of TL1A in serum of the DM 1 month group,the DM 3 month group and the DM 6 month group were significantly lower than that of the control group (F=77.405,P＜0.05).The concentration of serum TNF-α and IL-1β increased after DM model was established (F=3.508,15.416; P＜0.05); the VEGF level in serum showed no difference between the groups (F=1.242,P＞0.05).The vitreous TL1A levels of the control group,the DM 1 month group,the DM 3 month group and the DM 6 month group were (91.50±8.18),(67.03±6.74),(47.44±4.92) and (46.01±4.62) ng/L respectively,every DM groups showed significant difference with the control group (F=114.777,P＜0.05); VEGF level in vitreous increased from 1 month after DM model was established (F=8.816,P＜0.05) ; TNF-α and IL-1β level in vitreous also showed an upward tendency (F=4.392,3.635; P＜0.05).Paraffin section immunohistochemistry showed that the absorbance (also called optical density) of TLIA of the DM 1 month group and the DM 3 month group were significantly lower than that of the control group (t=6.851,6.066; P＜0.05),but the DM 6 month group showed no difference with the control group (t=1.401,P＞0.05) ; the level of VEGF and TNF-α in DM groups were higher than that of the control group (tVEGF =-4.709,-16.406,-9.228; tTNF-α =-4.703,-6.583,-17.762; P＜0.05) ; the level of IL-1β were significantly higher in the DM 1 month group and the DM 6 month group (t=-4.108,-3.495; P＞0.05); but the DM 3 month showed no difference with the control group (t=-0.997,P＞0.05).HE staining of retina showed that the retina of the control group and the DM 1 month group had normal retinal structures,the DM 3 month group had retinal edema and disorganization,the DM 6 month group had severe retinal edema,deep stain of ganglion cells,and more neovascularization in inner plexiform layer.Conclusion VEGI is involved in the pathogenesis of DR,and it might interacts with VEGF,TNF-α and IL-1β to affect the development of DR.