凉血活血药物对视网膜新生血管影响的实验研究

【作者】 马珊；

【导师】 王明芳；

【作者基本信息】 成都中医药大学 ， 中医眼科学， 2008， 博士

【摘要】 目的:探讨凉血活血药物对视网膜新生血管的干预及其作用机理。方法:应用MTT法检测不同浓度的凉血活血含药血清对血管内皮细胞ECV304增殖的影响,检测细胞上清液中VEGF含量。通过氧诱导C57BL/6幼鼠的方法建立视网膜新生血管的动物模型.随机分为正常对照组、模型对照组、凉血活血药物高、低剂量组。墨汁视网膜血管显影及整装铺片观察幼鼠视网膜血管的形态;计数视网膜切片中突破血管内界膜的内皮细胞核数;免疫组织化学检测HIF-1和NF-κB在视网膜中的表达情况;测量幼鼠尾长、体重、脑重,了解高氧对实验幼鼠的生长发育是否有影响。结果:凉血活血药物可抑制ECV304细胞的增殖,同时降低细胞上清液中VEGF的表达。氧诱导后,幼鼠血管内皮细胞核数目明显增加,HIF-1、NF-κB明显上调,与正常对照组比,差异有统计学意义(P＜0.05),提示视网膜新生血管动物模型成功;各治疗组血管内皮细胞核数目减少、HIF-1、NF-κB下调,与模型组比,差异有统计学意义(P＜0.05)。提示凉血活血药物对视网膜新生血管有抑制作用。幼鼠尾长、体重、脑重与正常对照组相比降低,差异有统计学意义(P＜0.05)。提示高氧对实验动物的生长发育有一定影响。结论:凉血活血药物可抑制ECV304细胞增殖,其机理是通过下调VEGF来实现。凉血活血药物在体内可抑制视网膜新生血管的形成,可能是通过下调与血管生成相关的NF-κB、HIF-1有关。因此,凉血活血药物对视网膜新生血管防治将为临床应用提供一定的实验依据。更多还原

【Abstract】 Purpos:to study the therapeutic liangxuehuoxue effect on the supression of retinal neovascularizationMethods:the ECV304cells were cultured with different concentrations of liangxuehuoxue,The proliferation rate of endothelial cells are assayed with MTT methods,the level of VEGF was tested.The animal model of oxygen-induced retinopathy was made by subjecting postnal mice to hyperoxic conditions taken retinal angiogram with ink.Quantified the degree of hyperoxia-induced neocascularization in serial paraffin cross sections by counting the number of vascular cell nuclei on vitreal side of the intemal membrane,the expression of HIF-1 and NF-κB in the oxygen-induced retina with immuhistochemistry were observed,also the weight、the weight of brain and length were measureded.Results:Liangxuehuoxue could inhibit the proliferation of ECV304 cells and the expression of VEGF.Afier the mouse were induced by oxygen,the endothelial cell nuclei increased and HIF-1、NF-κB went up obviously,The length、weight、and weight of brain got down.Liangxuehuoxue caused the endothelial cell nuclei reduced and HIF-1、NF-κB descended.Coneulsions:liangxuehuoxue inhibits the ECV304 cells proliferation and oxygen-induced intravitreal retinal angiogenesis by down-regulated HIF-1and NF-κB related to the angiogenesis.Therefore,the treatment of liangxuehuoxue will provide certain experiment basis for the clinical practice.更多还原