基于VEGF途径探究丹参素对大鼠视网膜静脉阻塞及对视网膜功能的改善作用机制  

Mechanism of Danshensu in retinal vein occlusion and retinal functions in rats via the VEGF pathway

作　　者：刘志强[1] 李亚坤[1] 白玫[1] 郭向东[1] 梁春利[1] LIU Zhiqiang;LI Yakun;BAI Mei;GUO Xiangdong;LIANG Chunli(Ophthalmology Department of the Second Affiliated Hospital of Hebei North University,Zhangjiakou 075100,China)

机构地区：[1]河北北方学院附属第二医院眼科,河北张家口075100

出　　处：《中国比较医学杂志》2023年第5期52-60,共9页Chinese Journal of Comparative Medicine

基　　金：张家口市重点研发计划项目(2021070D)。

摘　　要：目的探讨丹参素对视网膜静脉阻塞(RVO)大鼠的改善作用及可能机制。方法随机选择8只SPF级雄性SD大鼠作为对照组,剩余大鼠左眼利用孟加拉红联合激光光动力法诱导BRVO模型,成模大鼠随机分为模型组、丹参素低(15 mg/kg)、中(30 mg/kg)、高剂量组(60 mg/kg),每组各8只,对照组及模型组大鼠每日经尾静脉注射等量生理盐水,连续21 d。荧光素眼底血管造影(FFA)技术观察视网膜静脉结构;耳静脉微循环检测静脉血液流速;全视野视网膜电图(ffERG)评估视网膜功能;光学相干断层扫描(OCT)测量视网膜层厚度;HE染色观察视网膜组织学变化;RT-qPCR技术检测VEGF mRNA相对表达量;Western blot技术检测视网膜组织中缺氧诱导因子1α(HIF1α)、细胞外信号调节激酶1/2(ERK1/2)、p-ERK1/2、黏着斑激酶(FAK)、p-FAK、VEGF及色素上皮衍生因子(PEDF)蛋白表达情况。结果给药前,与对照组比较,模型大鼠视网膜静脉血流中断、远端血管迂曲、扩张,视网膜厚度增加,ffERG a波、b波振幅降低,耳静脉血液流速降低(P<0.05);给药后,与对照组比较,模型组及丹参素三个给药组大鼠视网膜静脉阻塞处再通,视网膜厚度、ffERG a波、b波振幅及耳静脉血液流速降低(P<0.05),视网膜存在不同程度损伤,模型组大鼠视网膜结构不完整,神经节细胞层(GCL)细胞数目减少,外核层(ONL)感光细胞层几乎丧失,视网膜组织中VEGF mRNA及蛋白和HIF1α、p-ERK1/2、p-FAK蛋白相对表达量升高,PEDF蛋白相对表达量降低(P<0.05);与模型组比较,丹参素三个给药组大鼠上述指标及因子表达水平均逆转(P<0.05),视网膜病理损伤逐渐减轻;丹参素作用效果呈剂量依赖性(P<0.05)。结论丹参素可减轻RVO大鼠视网膜组织损伤,促进阻塞静脉微循环恢复并改善视网膜功能,其作用机制可能与抑制VEGF血管生成相关途径的激活有关。Objective To investigate the ameliorative effect and possible mechanism of Danshensu in retinal vein occlusion(RVO)rats.Methods Eight SPF grade male SD rats were randomly selected as the control group.The RVO model was induced by Bengal red combined with the laser photodynamic method in the left eye of the remaining rats.The model rats were randomly divided into model,Danshensu low(15 mg/kg),medium(30 mg/kg),and high dose group(60 mg/kg),with eight rats in each group.Rats in control and model groups were injected with the same amount of normal saline through the caudal vein every day for 21 days.Fundus fluorescein angiography was used to observe the retinal vein structure.The venous blood flow velocity was measured by ear vein microcirculation.A full field electroretinogram(ffERG)was used to evaluate retinal functions.The retinal layer thickness was measured by optical coherence tomography.Histological changes of the retina were observed by HE staining.The relative expression of VEGF mRNA was measured by RT⁃qPCR.Expression of hypoxia inducible factor 1α(HIF1α),extracellular signal regulated kinase 1/2(ERK1/2),p⁃ERK1/2,focal adhesion kinase(FAK),p⁃FAK,VEGF,and pigment epithelium⁃derived factor(PEDF)in retinal tissue was detected by Western blot.Results Before administration,compared with the control group,the retinal vein blood flow of model rats was interrupted,distal blood vessels were tortuous and dilated,the retinal thickness was increased,the amplitudes of ffERG a and b waves were decreased,and the blood flow velocity of the ear vein was decreased(P<0.05).After administration,compared with the control group,retinal vein occlusion was recanalized,the retinal thickness,amplitudes of ffERG a and b waves,and blood flow velocity of the auricular vein were decreased(P<0.05),and the retina was damaged by various degrees.The retinal structure of the model group was incomplete,the number of ganglion cell layer cells was decreased,the photoreceptor cell layer of the outer nuclear layer was almost lost,and rel

关 键 词：丹参素 视网膜静脉阻塞 视网膜功能 血管内皮生长因子 

分 类 号：R-33[医药卫生]