黄芪多糖对人心脏微血管内皮细胞增殖及再灌注后内皮细胞与中性粒细胞黏附的影响  

Effects of astragalus polysaccharides on the proliferation of human cardiac microvascular endothelial cells adherent to polymorphonuclear neutrophils after reperfusion injury

作　　者：陈立新 朱海燕[2] 朱陵群[3] 谢连娣 林钦[4] 张颖[4] 

机构地区：[1]北京王府中西医结合医院心血管内科,北京市102209 [2]北京中医药大学中医内科学教育部重点实验室,北京市100700 [3]北京中医药大学东直门医院中医脑病研究室,北京市100700 [4]北京中医药大学,北京市100029

出　　处：《中国组织工程研究与临床康复》2007年第27期5382-5386,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金资助项目(30572391)~~

摘　　要：目的:观察黄芪多糖对人心脏微血管内皮细胞的增殖效应及其对再灌注损伤人心脏微血管内皮细胞与人外周血中性粒细胞黏附作用的影响。方法:实验于2006-07/12在北京中医药大学东直门医院中医内科重点实验室完成。①实验材料:原代人心脏微血管内皮细胞(ScienCell公司);黄芪多糖(惠州市东方植物保健科技有限公司)。②实验过程及评估:实验使用体外培养的人心脏微血管内皮细胞第4,5代细胞,采用倒置相差显微镜下观察细胞形态和生长情况;以四甲基偶氮唑盐法检测经不同浓度黄芪多糖处理24h,以及经黄芪多糖100,50,25mg/L分别处理4,6,8h后的增殖变化;以体外缺糖缺氧2h模拟体内缺血,复糖复氧6h模拟再灌注复制人心脏微血管内皮细胞再灌注损伤模型;虎红法测定经缺氧再复氧处理后的人心脏微血管内皮细胞与外周血中性粒细胞的黏附;免疫细胞化学法和图象定量分析系统观察细胞间黏附分子1的蛋白表达变化。结果:①人心脏微血管内皮细胞形态和生长情况:人心脏微血管内皮细胞复苏48～72h后细胞呈铺路石样生长,在15个细胞倍增周期内,其形态、生物、生理学特性稳定。②不同浓度黄芪多糖对人心脏微血管内皮细胞增殖的影响:黄芪多糖浓度≥5g/L时,对细胞生长具有抑制作用(P<0.05或0.01);浓度为2.5g/L时,对细胞的增殖无影响;浓度在1.25g/L～18.78mg/L时,可明显促进细胞的增殖(P<0.05或0.01)。③黄芪多糖处理不同时间对人心脏微血管内皮细胞增殖的影响:经黄芪多糖100,50,25mg/L3个剂量分别处理4,6,8h后,其增殖与正常对照组比无明显变化。④黄芪多糖对人心脏微血管内皮细胞与中性粒细胞间黏附作用及人心脏微血管内皮细胞细胞间黏附分子1蛋白表达的影响:黄芪多糖50mg/L可显著抑制再灌注损伤人心脏微血管内皮细胞与外周血中性粒细胞的黏附(P<0.01);100mg/L可明显降低人�AIM： To observe the effects of astragalus polysaccharides （APS） on the proliferation of human cardiac microvascular endothelial cells （HCMEC） and adhesion between HCMEC injured by reperfusion and polymorphonuclear neutrophils （PMN）. METHODS： The experiment was performed at the Key Laboratory of Chinese Internal Medicine of Ministry of Education, Beijing University of Chinese Medicine from July to December 2006. The fourth and fifth generation HCMECs cultured in vitro were selected, and their morphous and growth was observed under inverted phase contrast microscope; the proliferation of HCMECs was detected by means of MTT after HCMEC was treated with APS of different concentrations for 24 hours and separately treated with 100, 50, and 25 μg/mL APS for 4, 6, and 8 hours. Ischemia/reperfusion （I/R） injury model was established by oxygen-glucose deprivation for 2 hours and recovery 6 hours method. The effect of APS on PMN adhesion to HCMEC after reperfusion injury was measured by rose Bengal staining. Expression of intracellular adhesion molecule-1 （ICAM-1） was determined by immunocytochemistry and image quantitative analysis system. RESULTS： （1）The HCMEC showed a typical cobblestone appearance after recovered for 48 to 72 hours and its morphological, biological and physiological characters were stable during the 15 population doubling periods. （2）The proliferation of HCMEC was lower （P〈 0.05 or 0.01） when APS was more than 5 g/L, and higher （P〈 0.05 or 0.01） when APS was between 1.25 g/L-18.78 mg/L. There was no obvious change in proliferation when treated with 2.5 g/L APS. （3）Compared with control group, there was no evident change in the proliferation of HCMEC when APS was treated separately with 100, 50, and 25 mg/L for 4, 6, and 8 hours. （4）50 mg/L APS could decrease the adhesion of PMN to HCMEC apparently （P 〈 0.01 ） and 100 mg/L of APS could reduce the expression of ICAM-1 in HCMEC significantly （P 〈 0.05）. CONCLUSION： （1）APS can promote

关 键 词：黄芪多糖 微血管内皮细胞 胞间粘附分子1 缺氧/复氧 组织构建 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]