健脾祛瘀法对血脂异常ApoE基因缺失小鼠视网膜RPE黑色素和脂褐素的影响  

Influence of invigorating spleen and removing blood stasis method on RPE melanin and lipofuscin of retinal on ApoE dyslipidemia gene deletion mouse

作　　者：王毅[1] 李娟[1] 李进辉[1] 李罗翔[1] 曾庆华[1] 

机构地区：[1]成都中医药大学附属医院,成都610072

出　　处：《中国中医基础医学杂志》2014年第1期54-57,76,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金资助项目-健脾祛瘀法调控AMD脉络膜血管内皮VEGF/VEGFR2通路的研究(30973774)

摘　　要：目的:观察健脾祛瘀法对血脂异常ApoE基因缺失(apolipoprotein E-deficient,ApoE-/-)小鼠视网膜色素上皮细胞(retinal pigment epithelium,RPE)黑色素和脂褐素的影响。方法:随机将60只2月龄ApoE-/-小鼠分为普食组、高脂组、高剂量治疗组、中剂量治疗组、低剂量治疗组,喂养5个月后检测其血浆总胆固醇(total plasma cholesterol,TC)、低密度脂蛋白(low density lipoprotein,LDL)、血浆甘油三酯(plasma triglyceride,TG)含量,银氨液浸染法和醛品红法染色后光镜下观察视网膜色素上皮细胞(RPE)形态结构,并半定量检测RPE细胞内黑色素和脂褐素含量及计算其黑色素/脂褐素比值,采用多元线性回归分析黑色素、脂褐素、黑色素/脂褐素比值与TC、LDL、TG相关性。结果:TC、TG和7月龄ApoE-/-各治疗组小鼠比7月龄ApoE-/-高脂组明显降低(P<0.001),LDL和7月龄ApoE-/-高、中治疗组小鼠比7月龄ApoE-/-高脂组明显降低(P<0.001),7月龄ApoE-/-低剂量治疗组比7月龄ApoE-/-高脂组降低(P>0.05)。光镜下,7月龄ApoE-/-高脂组和7月龄ApoE-/-普食组小鼠RPE排列紊乱、细胞萎缩,7月龄ApoE-/-各治疗组小鼠视网膜RPE排列较整齐、细胞萎缩减轻。半定量检测结果,与7月龄ApoE-/-高脂组比,7月龄ApoE-/-各治疗组小鼠视网膜RPE细胞内黑色素含量明显增多(P<0.001),7月龄ApoE-/-各治疗组小鼠视网膜RPE细胞内脂褐素含量明显降低(P<0.001),7月龄ApoE-/-各治疗组小鼠视网膜RPE细胞内黑色素/脂褐素比值明显增多(P<0.001)。采用逐步引入-剔除法进行多元线性回归分析发现,黑色素含量与TC、LDL、TG呈负直线相关,回归方程为:黑色素含量=1837732.32-63225.96×胆固醇,脂褐素含量与TC、TG呈正直线相关,回归方程为:脂褐素含量=418363.52+139394.42×胆固醇,黑色素/脂褐素比值与TC、TG呈负直线相关,回归方程为:黑色素/脂褐素=2.057-0.127×胆固醇。结论:健脾祛瘀法可通过降低血脂异常ApoE-/-小鼠高脂Objectives： To observe effect of tonifying Spleen and promoting blood circulation on the melanin and the lipofuscin of retinal pigment epithelium （RPE） in apolipoprotein E-deficiency （ApoE-/-） mice with dyslipidemia. Methods： sixty apolipoprotein E-deficiency （ApoE-/-） mice at two-month age were randomly divided into normal diet group, high fat diet group, and high dose experimental group and middle dose experimental group and low dose experimental group. After being fed for 5 months, plasma total cholesterol （TC）, low density lipoprotein （LDL） and triglyceride （TG） were determined, meanwhile, morphological changes of retinal pigment epithelial cells were observed under light microscope with Fontana staining and aldehydo-magenta staining, and amounts of melanin and lipofuscin of retinal pigment epithelial cells and ratio of the melanin and the lipofuscin were counted by semi-quantitative histopathology,and correlation of the melanin and the lipofuscin and ratio of the melanin and the lipofuscin with TC, LDL and TG were analysed with muhinomial linear regression. Results： Compared with high fat diet group, plasma TC and TG were significantly decreased in every experimental group （P 〈 0. 001 ） , plasma LDL were significantly decreased in high dose experimental group and middle dose experimental group （ P 〈 0. 001 ） , and plasma LDL were decreased in low dose experimental group （ P 〉 0.05 ）. Under light microscope, RPE were disorganized and atrophic in both high fat diet group and normal diet control group, However, in every experimental group, RPE were well organized, less atrophy. By semi-quantitative histopathology,Compared with high fat diet group,melanin of retinal pigment epithelial cells were significantly increased in every experimental group （P 〈 0. 001 ） , the lipofuscin were significantly decreased in every experimental group （P 〈 0. 001 ） ,the ratio of the melanin and the lipofuscin were significantly increased in every experimental group （ P

关 键 词：血脂异常 ApoE- -小鼠 色素上皮细胞 黑色素 脂褐素 健脾祛瘀法 年龄相关性黄斑变性 

分 类 号：R285.5[医药卫生—中药学]