地黄低聚糖对人脂肪组织源性间充质干细胞分泌血管内皮细胞生长因子的影响  

The effect of rehmannia glutinosa oligosaccharide on vascular endothelial cell growth factor secreted by human adipose tissue-derived mesenchymal stem cells

作　　者：王玉红[1,2] 陈光辉[1] 王舒[1] 张琰琴[1] 王磊[1] 秦宇红[3] 

机构地区：[1]解放军总医院心血管内科,北京100853 [2]北京军区总医院急诊科,北京100700 [3]解放军总医院第一附属医院急诊科,北京100037

出　　处：《心脏杂志》2009年第3期332-335,346,共5页Chinese Heart Journal

基　　金：国家自然科学基金项目资助(30471924/30572445)

摘　　要：目的探讨地黄低聚糖(RGO)对分离、培养的人脂肪组织源性间充质干细胞(ADMSCs)分泌血管内皮细胞生长因子(VEGF)的影响。方法人脂肪组织用2.5 g/L胶原蛋白酶Ⅰ消化、分离ADMSCs。取沉淀的细胞进行培养,用免疫细胞化学染色法鉴定其表面CD44、CD105、CD45和CD34分子的表达。以IMDM培养基作为空白对照,加入RGO配成不同浓度(0、1、10、100及400 mg/L)的IMDM分别培养ADMSCs,用MTT比色法检测ADMSCs的增殖。培养72 h后,用ELISA法测定不同培养基中VEGF的含量。结果免疫细胞化学染色显示,分离培养的细胞表面CD44+、CD105+、CD34-、CD45-。MTT比色法的结果显示,与空白对照组相比,1和10 mg/L组无明显差别,100及400 mg/L组明显升高(均P<0.01),且100 mg/L组明显高于400 mg/L组(P<0.05)。0、1、10、100、400 mg/L5个组VEGF的含量,分别为(627.88±33.86)、(655.50±40.29)、(666.50±43.20)、(843.50±53.05)和(722.88±51.34)ng/L。结论RGO对ADMSCs的增殖有促进作用,且呈一定的浓度依赖性,并导致其分泌VEGF的作用增强。AIM To isolate and culture human adipose tissue-derived mesenchymal stromal cells （hADMSCs） and to study the effect of rehmannia glutinosa oligosaccharide （RGO） on the secretion of vascular endothelial cell growth factor（VEGF） from ADMSCs. METHODS Human adipose tissue from patients receiving abdominal surgery was digested with 2.5 g/L collagenase type I , hADMSCs were separated and cells from sediment were cultured. The expression of CD44, CD105, CD45 and CD34 on ADMSCs were examined by immunocytochemical staining. ADMSCs were cultured in IMDM culture medium containing different concentrations （0,1,10,100,400mg/L） of RGO. The effects of different concentrations of RGO on hADMSCs proliferation were measured by MTT colorimetry. At 72 hours after culture, ELISA was used to detect the level of VEGF in different culture mediums. RESULTS ADMSCs were identified by positive expression of CD44 and CD105 and negative expression of CD34 or CD45.According to MTr colorimetry＇s results, there was little and insignificant difference in proliferation between groups of 1 mg/L and 10 mg/L and control group. Significant proliferation was seen in ADMSCs of groups of 100 mg/L and 400 mg/L （P 〈 0.01 ）. The contents of VEGF were （627.88± 33.86）, （655.50 ±40.29）, （666.50±43.20）, （843.50 ±53.05） and （722. 88 ±51.34） ng/L, respectively. CONCLUSION RGO has significant effect on ADMSCs＇s proliferation and can promote secretion of VEGF.

关 键 词：脂肪组织源性间充质干细胞 血管内皮生长因子 地黄低聚糖 

分 类 号：R282.71[医药卫生—中药学]