【作者】 马栋；

【导师】 郭承伟；

【作者基本信息】 山东中医药大学 ， 中医五官科学， 2013， 博士

【摘要】 目的：针对糖尿病视网膜病变（DR）虚劳络阻的病机特点，以络病为切入点，评价络刺结合药物通络对DR微血管损伤的修复作用。方法：以大鼠作为实验研究对象，对其注射链佐脲菌素（STZ）进行DR造模，采用络刺加大黄蛰虫丸灌胃给药，治疗后观察以下指标：1.大鼠的体重、血糖、糖化血红蛋白及血浆纤维蛋白原，并观察白内障的发生率。2.眼底荧光血管造影（FFA）观察DR大鼠眼底血管管径、荧光素渗漏形态及面积的变化。3.视网膜组织石蜡切片结合HE染色，光镜下观察视网膜各层组织结构的变化。4.视网膜组织石蜡切片联合免疫组织化学法，观察视网膜中核因子-κb（nf-κb）的表达。5.视网膜消化铺片联合PAS染色，观察对视网膜微血管形态学上的改变。6.视网膜消化铺片结合免疫荧光染色技术，观察视网膜血管壁中血管细胞粘附分子-1（VCAM-1）的表达。结果：1.络治能明显改善糖尿病大鼠三多一少等症状，减轻血糖的增高对大鼠体重的影响，降低血液中纤维蛋白原含量。2.FFA显示，络治能减少DR大鼠眼底新生血管的生成，减轻血管渗漏。3.视网膜组织切片形态学观察结果显示，络治能明显减轻视网膜组织水肿程度，减轻神经细胞损伤，抑制新生血管生成。4.免疫组织化学染色显示，络治可下调视网膜中nf-κb的表达。5.视网膜消化铺片联合PAS染色及免疫荧光染色显示络治能改善微血管网形态，并下调血管壁中炎性因子VCAM-1的表达。结论：络治法能够有效减轻高血糖引起的视网膜微血管损伤，改善糖尿病视网膜微血管的结构与机能，改善微血管管壁的通透性，抑制或消除诱发的出血、渗出，实现对视网膜保护作用。其作用机理可能是通过调节血浆中纤维蛋白原的含量，降低了血液粘稠度，增加局部视网膜血流量及氧供，抑制视网膜中nf-κb的活化及下游VCAM-1等炎性因子的表达等方面得以实现。上述研究成果在丰富中医络脉理论的同时，为DR的防治提供了新的思路与方法。更多还原

【Abstract】 Objective: For diabetic retinopathy (DR) characteristics of consumptive diseasepathogenesis of collaterals, by collateral disease as the breakthrough point to evaluate thethe role in repairment of diabetic retinal microvascular injury treated with the collateralneedling and drug。 Methods: Using rats as experimental subjects， by injection ofstreptozocin (STZ) into DR model，after treatment with Dahuang Zhechong pillsintragastric administration and collateral puncture，in observation of the followingindicators:1.Rat body weight、blood glucose、glycosylated haemoglobin、plasmafibrinogen and the incidence of cataracts.2. By fundus fluorescein angiography (FFA), toobserve the changes of retinal vessel diameter of DR rats, fluorescein leakage area andshape.3.By paraffin section of retinal tissue combined with HE staining method,to observethe change of every layer of retina tissue structure under light microscope.4.By paraffinsection of retinal tissue combined with immunohistochemical method,to observe theexpression of retinal nuclear factor-κb (nf-κb)。5. Through the retinal digest preparationscombined with PAS staining, to observe retinal microvascular changes inmorphology.6.Through the retinal digest preparations combined with immunofluorescencetechnique, to observe the expression of vascular cell adhesion molecule-1(VCAM-1) inretinal vascular wall.Results:1.Collaterals therapy can significantly change the symptomsof“three more and one less”in DR rats, reduce the effect of high glucose on rat bodyweight,and reduce blood fibrinogen content.2.FFA showed that collaterals therapy canreduce the generation of DR rat retinal neovascularization，and can reduce vascularleakage.3. Morphological observation of retinal tissue biopsies showed that collateralstherapy can significantly reduce the degree of retinal edema, reducing neuronal damage,inhibit tumor angiogenesis.4.Immunohistochemistry showed collaterals treatmentdecreased expression of nf-κB in retina.5.Retinal digest preparations combined with PAS and immunofluorescence showed collaterals therapy can improve the microvascularmorphology, and reduce the expression of inflammatory factor VCAM-1in the vascularwall.Conclusion: Collateral treatment can effectively alleviate the hyperglycemia inducedby retinal microvascular injury, improve diabetic retinal structure and function of bloodvessels, change the microvessel wall permeability, inhibit or eliminate induced hemorrhageand exudation, thus realizing the retinal protection. Its mechanism may be through theregulation of fibrinogen in plasma content, reducing the blood viscosity, increase the flowand oxygen partial retinal blood supply, inhibit the expression of retinal nf-κB activationand downstream of the VCAM-1and other inflammatory factors. The above researchresults in the rich of traditional Chinese collaterals theory,and at the same time providesnew ideas and methods for the prevention and treatment of DR.更多还原