文献类型：期刊文献

中文题名：低氧环境下黄芪甲苷促进人骨髓间充质干细胞分泌血管内皮细胞相关因子的实验研究

英文题名：Astragaloside Promoted Human Bone Marrow Mesenchymal Stem Cells to Secrete Cytokine Related Vascular Endothelial Cells under Hypoxia Condition

作者：胡继宏[1];贾佳[1,2];路娟[1];王秋萍[1];赵静苗[1];金华[1];靳利梅[1];赵翊[1];李金娟[1]

第一作者：胡继宏

机构：[1]甘肃中医药大学科研实验中心,兰州730000;[2]西安海棠职业学院中医教研室,西安713000

第一机构：甘肃中医药大学科研实验中心（甘肃省中医药标准化技术委员会秘书处）

年份：2018

卷号：38

期号：6

起止页码：693

中文期刊名：中国中西医结合杂志

外文期刊名：Chinese Journal of Integrated Traditional and Western Medicine

收录：CSTPCD;;Scopus;北大核心:【北大核心2017】；CSCD:【CSCD2017_2018】；PubMed;

基金：国家自然科学基金资助项目(No.81260597);甘肃省高校基本科研业务费项目(No.2012-3)

语种：中文

中文关键词：血管内皮生长因子;黄芪甲苷;低氧;骨髓间充质干细胞;内皮素;前列环素;内皮型一氧化氮合酶

外文关键词：vascular endothelial growth factor;astragaloside;hypoxia;bone mesenchymal stem cell;endothelin;prostacyclin;endothelial nitric oxide synthase

摘要：目的观察在低氧环境下,黄芪甲苷能否诱导人骨髓间充质干细胞(BMSCs)向血管内皮样细胞分化,提高BMSCs经血管内皮生长因子(VEGF)诱导分化后的血管内皮样细胞的功能。方法将人BMSCs分为4组:对照组(单纯人BMSCs培养)、VEGF组(含VEGF载体的腺病毒的完全培养基加入人BMSCs)、黄芪甲苷组(含黄芪甲苷粉剂加入人BMSCs)和联合诱导组(含黄芪甲苷粉剂和VEGF载体的腺病毒的完全培养基加入人BMSCs),5%O2干预2周,对分化的细胞进行形态学观察,流式细胞仪检测细胞表型CD31和CD105的表达,ELISA检测细胞培养上清液中VEGF及内皮型一氧化氮合酶(eNOS)因子的含量,Real-time PCR和Western blot检测内皮素(ET)和前列环素(PGI)的表达。结果 (1)2周后,镜下观察对照组与黄芪甲苷组细胞形态无明显变化,VEGF组和联合诱导组细胞似铺路石子样生长。(2)流式结果显示,对照组与黄芪甲苷组细胞不表达CD31,高表达CD105;VEGF组和联合诱导组细胞高表达CD31,低表达CD105,且VEGF组CD31表达高于联合诱导组(P<0.05),而CD105低于联合诱导组(P<0.05)。(3)ELISA检测发现,与对照组比较,各干预组的VEGF及eNOS因子表达均增高(均P<0.05),且大小顺序为VEGF组>联合诱导组>黄芪甲苷组。(4)RT-PCR及Western blot检测发现,与对照组比较,各干预组ET水平均升高(P<0.05),PGI表达量均降低(P<0.05);且联合诱导组ET蛋白表达水平最高,VEGF组PGI表达量最低。结论在低氧环境下,黄芪甲苷促进人BMSCs分泌VEGF相关因子;黄芪甲苷联合VEGF可以促进VEGF诱导分化后的血管内皮样细胞分泌ET。
Objective To observe whether astragaloside could induce human bone marrow mesenchymal stem cells（ BMSCs） to differentiate into vascular endothelial cells（ VECs）,or improve the function of VECs under hypoxia condition.Methods The third passage BMSCs were divided into four groups,i. e.,the blank control group（ cultured with conventional culture medium）,the VEGF induction group [cultured with adenovirus vector containing vascular endothelial growth factor（ VEGF） ],the Astragaloside group（ cultured with astragaloside）,and the combined group（ cultured with adenovirus vector containing VEGF and astragaloside）. After intervened by 5% O2 for 2 weeks,morphologies of differentiated cells were observed. The expressions of cell phenotype CD31 and CD105 were detected by flow cytometry. The levels of VEGF and endothelial nitric oxide synthase（ eNOS） in supernate were detected by ELISA. The expressions of endothelin（ ET） and prostacy-clin（ PGI） were detected by Real-time PCR（ RT-PCR） and Western blot. Results（ 1） No obvious changes occurred in the cell morphology in the blank control group and the Astragaloside group after two weeks. Cells in the VEGF group and the combined group exhibited a colony-like growth.（ 2） Results of flow cytometry showed that CD31 was negatively expressed in the blank control group and the Astragaloside group,but the expression of CD105 was high. However,the expression of CD31 increased,and the expression of CD105 decreased in the VEGF group and the combined group. The positive rate of CD31 was significantly higher in the VEGF group than in the combined group（P 〈 0. 05）,and the positive rate of CD105 expression was lower in the VEGF group than in the combined group（P 〈 0. 05）.（ 3） ELISA showed,as compared with the blank control group,the expressions of VEGF and eNOS increased significantly in all the intervention groups（P 〈 0. 05）,ranked as the VEGF group 〉 the combined group 〉 the Astragaloside group.（ 4） RT-PCR and Western blot showed,as compared with the blank control group,ET level increased（P 〈 0. 05） and PGI decreased（P 〈 0. 05） in all the intervention groups. Further more,the expression of ET was the highest in the combined group,and the expression of PGI was the lowest in the VEGF group. Conclusions Astragaloside promoted BMSCs to secrete VEGF related factors under hypoxia condition.Astragaloside combined VEGF could promote the secretion of ET by VECs after VEGF induced differentiation.