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目的观察鼓槌石斛乙醇提取物(ethanol extract of Dendrobium chrysotoxum,EEDC)对链脲佐菌素(STZ)诱导糖尿病小鼠非增殖性糖尿病视网膜病变(NPDR)的改善作用及其机制。方法采用STZ诱导小鼠发生糖尿病,进而建立糖尿病并发症NPDR实验动物模型,再给予EEDC进行干预。采用视网膜组织伊文思蓝渗漏实验观察NPDR小鼠中血-视网膜屏障(BRB)的破坏情况;采用real-time PCR(RT-PCR)检测视网膜组织中白细胞介素-6(IL-6)、IL-1β、肿瘤坏死因子(TNF-α)及早期生长反应因子(Egr-1)、组织因子(TF)、纤溶酶原激活物抑制剂1(Serpine 1)的基因表达;进一步采用酶联免疫实验(ELISA)检测血清中IL-1β、IL-6、TNF-α及TF、Serpine 1的量。结果与对照组相比,模型组小鼠视网膜组织伊文思蓝渗漏量明显增加,EEDC能够减少视网膜组织伊文思蓝渗透量;EEDC能够显著降低模型小鼠视网膜组织中增加的IL-1β、IL-6、TNF-α及Egr-1、TF、Serpine 1的基因表达;EEDC能够明显下调模型小鼠血清中增加的IL-1β、IL-6、TNF-α及TF、Serpine 1的量。结论 EEDC具有改善STZ诱导小鼠NPDR的作用,其机制为通过抑制NPDR小鼠视网膜和血清中增加的炎性因子IL-1β、IL-6、TNF-α以及与凝血/纤溶相关基因TF、Serpine 1的基因表达,缓解BRB的破坏,从而发挥改善NPDR的作用。
Objective To investigate the effect and mechanism of ethanol extract of Dendrobium chrysotoxum (EEDC) on non-proliferative diabetic retinopathy (NPDR) induced by streptozotocin (STZ) in diabetic mice. Methods STZ was used to induce diabetes in mice and then NPDR experimental animal model of diabetic complications was established, and then EEDC was given for intervention. The damage of blood-retinal barrier (BRB) in NPDR mice was observed by Evans blue leak test of retina tissue. The levels of interleukin-6 (IL-6) IL-1β, TNF-α, Egr-1, TF and Serpine 1 were detected by enzyme linked immunosorbent assay Serum levels of IL-1β, IL-6, TNF-α and TF, Serpine 1 were measured by ELISA. Results Compared with the control group, the leakage of Evans blue in retinal tissue of mice in the model group was significantly increased, and EEDC reduced the amount of Evans blue permeation in the retinal tissue. EEDC significantly decreased the levels of IL-1β, IL -6, TNF-α, Egr-1, TF and Serpine 1. EEDC significantly decreased the levels of IL-1β, IL-6, TNF-α, TF and Serpine 1 in serum of model mice. Conclusion EEDC can improve the neuroprotective effect of STZ on NPDR in mice by inhibiting the increase of inflammatory cytokines IL-1β, IL-6, TNF-α in the retina and serum of NPDR mice as well as the effects of coagulation / fibrinolysis related gene TF, Serpine 1 gene expression, relieve the destruction of BRB, which play to improve the role of NPDR.