健脾祛瘀法对血脂异常ApoE基因缺失小鼠视网膜血管瘤增殖的影响  

Effect of tonifying spleen and promoting blood circulation on retina angiomatous proliferation in apolipoprotein E-deficiency mouse with dyslipidemia

作　　者：王毅[1] 庄利萍 李罗翔[1] 李娟[1] 曾庆华[1] 莫亚[1] 刘小虎[1] 

机构地区：[1]成都中医药大学附属医院,四川成都610072 [2]成都市都江堰中医院,四川成都611830

出　　处：《时珍国医国药》2013年第12期3046-3050,共5页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.30973774)

摘　　要：目的观察健脾祛瘀中药制剂对血脂异常ApoE基因缺失(ApoE-/-)小鼠视网膜血管瘤增殖的治疗作用。方法采用随机数字表法根据小鼠体质量分层随机分组。将36只2月龄ApoE-/-小鼠分为普通饮食组、高脂模型组及健脾祛瘀治疗组。高脂模型组及健脾祛瘀治疗组小鼠均用高脂饲料喂养,健脾祛瘀治疗组小鼠于实验的最后1个月给予健脾祛瘀制剂灌胃,每次0.3ml,每日2次,而普通饮食组和高脂模型组小鼠用等量生理盐水灌胃。于小鼠7月龄以过量麻醉法处死后制备左眼视网膜组织切片行免疫组化(SP法)染色血管内皮生长因子(VEGF)、血管内皮生长因子受体-2(VEGFR-2),光学显微镜观察并采用Mias 2000图像分析系统半定量检测3组动物视网膜色素上皮细胞(RPEC)内VEGF、视网膜外丛状层(OPL)内VEGFR-2含量、OPL微血管密度(MVD)及微血管腔面积(MVA),提取右眼视网膜组织行蛋白免疫印迹法(Western-Blot)检测3组动物视网膜VEGF蛋白表达。结果 7月龄健脾祛瘀治疗组小鼠视网膜OPL微血管密度、微血管腔面积均比同龄高脂模型组和普通饮食组明显减少(P<0.01-0.001),7月龄健脾祛瘀治疗组小鼠RPE细胞VEGF面积、OPL内VEGFR-2面积均比同龄高脂模型组和普通饮食组显著降低(P<0.05-0.01),7月龄健脾祛瘀治疗组小鼠视网膜VEGF/actin蛋白相对表达量比同龄高脂模型组明显降低(P<0.05),比普通饮食组高(P>0.05)。结论健脾祛瘀中药制剂可降低血脂异常ApoE-/-小鼠视网膜VEGF蛋白表达、RPE细胞内VEGF含量及视网膜外丛状层VEGFR-2含量,使视网膜外丛状层微血管密度降低、微血管腔面积缩小,抑制视网膜血管瘤增殖,对新生血管性年龄相关性黄斑变性有治疗作用。Objective To observe the effect of tonifying spleen and promoting blood circulation on retina angiomatous proliferation in apolipoprotein E - deficiency （ ApoE - / - ） mice with dyslipidemia. Methods Thirty - six ApoE - / - mice aged 2 - month - old were randomly divided into normal diet group, high fat diet group and medicine group. Higher content of fat diet was given for consecutive 5 months in the mice of the high fat diet group and medicine group, and in the last month, tonifying spleen and promo- ting blood circulation was administered by intragastric injection in the medicine group, and the equivalent normal saline solution was used in the same way in the normal diet group and high fat diet group. The numbers of vascular endothelial growth factor （VEGF） in retinal pigment epithelial（RPE） cells, vascular endothelial growth factor receptor - 2 （ VEGFR - 2） in outer plexiform layer（OPL） and microvascular density （MVD）and microvaseular area（MVA） in OPL were examined with immunohistochemistry staining sections of the miceg left eye by semi - quantitative histopathology with Mias 2000 Imaging Analyzer System. The numbers of retinal VEGF were examined with retinal of the mice~ right eye by Western - Blot. Results The MVD and MVA in retinal OPL in 7 - month - old mice of the medicine group, showing a significant reduce in comparison with the high fat diet group and in the normal diet group（ P 〈 0.01 - 0. 001 ）. The numbers of VEGF in RPE cells and the numbers of VEGFR - 2 in retinal OPL in 7 - month - old mice of the medicine group, showing a significant reduce in comparison with the high fat diet group and in the normal diet group（P 〈0.05 -0.01 ）. The relative numbers of retinal VEGF/actin were in 7 -month- old mice of the medicine group showing a significant reduce in comparison with in high fat diet group（ P 〈 0.05 ） , and showing a increase in normal diet control group（P 〉 0.05 ）. Conclusion Tonifying spleen and promoting blood circulation can reduce the numb

关 键 词：血脂异常 ApoE- -小鼠 视网膜 外丛状层、视网膜色素上皮 血管瘤增殖 年龄相关性黄斑变性 传统中医药 健脾祛瘀中药颗粒 

分 类 号：R285.5[医药卫生—中药学]