血管紧张素Ⅱ对骨髓基质干细胞源性成骨细胞血管内皮生长因子表达的影响  

Effect of angiotensin Ⅱ on the expression of vascular endothelial growth factor in human bone marrow stromal cells derived osteoblasts

作　　者：苏万汉[1] 陈滨[1] 方锦涛[1] 赵培冉[1] 

机构地区：[1]南方医科大学南方医院创伤骨科,广州510515

出　　处：《中华创伤骨科杂志》2012年第10期889-892,共4页Chinese Journal of Orthopaedic Trauma

基　　金：广东省自然科学基金课题（9151051501000075）;教育部博士点项目（20094433120016）

摘　　要：目的探讨血管紧张素Ⅱ对人骨髓基质干细胞（BMSCs）源性成骨细胞血管内皮生长因子（VEGF）基因表达及蛋白分泌的影响。方法分离、培养人BMSCs，并向成骨细胞方向诱导，用碱性磷酸酶（ALP）染色鉴定成骨细胞。取诱导14d的人BMSCs，分成5组（n=3），分别用0、50、100、150、200nmol／L血管紧张素Ⅱ处理细胞，以确定最佳作用浓度。取诱导21d的人BMSCs，分成5组（n=6），分别用最佳作用浓度血管紧张素Ⅱ处理细胞0、6、12、24、48h，采用实时定量逆转录．聚合酶链反应（RT．PCR）检测不同时间组VEGFmRNA的表达，采用酶联免疫吸附测定法（ELISA）检测不同时间组VEGF蛋白的表达。结果人BMSCs经诱导后ALP染色大部分呈阳性反应。0、50、100、150、200nmol／L血管紧张素Ⅱ处理的BMSCsVEGFmRNA表达平均分别为1．000．4-0．000、2．304±0．315、4．336±0．400、5．573±0．608、5．492±0．460，其中0、50nmol／L浓度组分别与100、150、200nmol／L浓度组比较差异均有统计学意义（P〈0．05）。选取150nmol／L为血管紧张素Ⅱ的最佳作用浓度。0、6、12、24、48h组细胞VEGFmRNA表达平均分别为1．000±0．000、1．984±0．160、4．372±0．378、5．573±0．586、4．994±0．445，其中0、6h组分别与12、24、48h组比较差异均有统计学意义（P〈0．05）。0、6、12、24、48h组细胞VEGF蛋白分泌量逐渐增加，且呈时间依赖性。结论用血管紧张素Ⅱ处理人BMSCs源性成骨细胞能够使VEGFmRNA表达量和蛋白分泌量快速增加。Objective To study the effect of Angiotensin H （Ang N） on the expression and protein secretion of vascular endothelial growth factor （VEGF） in human bone marrow stromal cells （hBMSCs） derived osteoblasts. Methods hBMSCs were isolated, cultured and induced to differentiate into osteoblasts. The cells were identified by alkaline phosphatase （ALP） staining. The hBMSCs induced for 14 days were divided into 5 groups （ n = 3） and treated by Ang Ⅱ at concentrations of 50, 100, 150 and 200 nmol/L to determine the optimal one. The hBMSCs induced for21 days were divided into 5 groups （n = 6） and treated by Ang Ⅱ at the optimal concentration for 0, 6, 12, 24, 48 hours. The level of mRNA VEGF was determined by SYBRGREN fluorescent quantitation and the level of VEGF protein in the medium was measured by ELISA. Results When the hBMSCs were cultured in the conditioned culture medium, most of the ALP staining was positive. The expressions of mRNA VEGF in the hBMSCs treated by Ang Ⅱ at concentrations of 0, 50, 100, 150 and 200 nmol/L were respectively 1.000 ±0.000, 2. 304±0. 315, 4. 336 ±0.400, 5. 573± 0. 608 and 5. 492 ± 0. 460. There were significant differences regarding the expression levels re- spectively between the first 2 concentrations and the latter 3 concentrations （ P 〈 0.05）. The concentration of 150 nmol/L was used as the optimal one to treat AngⅡ. The expressions of mRNA VEGF in the hBMSCs treated by Ang Ⅱ at the optimal concentration for 0, 6, 12, 24, 48 hours were respectively 1. 000 ± 0. 000, 1. 984 ± 0. 160, 4. 372 ±0. 378, 5. 573 ± 0. 586 and 4. 994 ± 0. 445. There were significant differences regarding the expression levels respectively between the first 2 time points and the latter 3 time points （ P 〈0.05）. The protein secretions by VEGF induced by Ang Ⅱ at the optimal concentration for 0, 6, 12, 24, 48 hours were increased in a time-dependent manner. Conclusion Incubation of hBMSCs derived osteoblasts with Ang Ⅱ may induce a rapid increase in both

关 键 词：间质干细胞 血管紧张素Ⅱ 血管内皮生长因子A 基因表达 

分 类 号：R735.2[医药卫生—肿瘤]