胰岛素对脂肪干细胞生长因子分泌功能的影响  

Effects of insulin on the growth factor secreting function of adipose-derived stem cells

作　　者：折涛[1] 胡大海[1] 张军[1] 刘佳琦[1] 张万福[1] 蔡维霞[1] 赵周婷[1] 汤朝武[1] 

机构地区：[1]第四军医大学西京医院全军烧伤中心,西安710032

出　　处：《中华烧伤杂志》2009年第4期268-271,共4页Chinese Journal of Burns

基　　金：国家自然科学基金（30772249）

摘　　要：目的了解不同浓度胰岛素对脂肪于细胞（ADSC）分泌生长因子功能的影响。方法获取人腹部脂肪组织分离培养ADSC，选择第3代细胞经免疫表型鉴定及脂源性诱导分化鉴定。根据培养液中胰岛素浓度，将ADSC分为1×10^-8、1×10^-7、1×10^-6mol／L3个实验组，常规培养至70％融合时，更换为含不同浓度胰岛素的无血清DMEM培养液培养3d。设无胰岛素培养液培养的ADSC为对照组。采用酶联免疫吸附测定法，测定ADSC的血管内皮生长闪子（VEGF）、肝细胞生长因子（HGF）分泌量；应用噻唑蓝比色法和羟脯氨酸比色法，检测ADSC上清液对Fb增殖及胶原合成的影响。结果1×10^-8、1×10^-7mol／L胰岛素组ADSC的VEGF、HGF分泌量分别为（471±41）、（762±66）ng／L和（643±64）、（930±67）ng／L，与对照组（286±47）、（577±84）ng／L比较，其含量显著增加（P〈0．05或P〈0．01）；而1×10^-6mol／L胰岛素组ADSC的VEGF、HGF分泌量无明显变化（P〉0．05）。1×10^-8、1×10^-7mol／L胰岛素组ADSC培养上清液有明显的促Fb增殖及胶原合成作用，以1×10^-7mol／L组作用更显著（P〈0．05或P〈0．01）。结论1×10^-8、1×10^-7mol／L胰岛素能显著促进ADSC分泌VEGF和HGF。Objective To study the effect of insulin in different concentrations on secretion function of growth factors of adipose-derived stem cells （ADSCs）. Methods ADSCs were isolated from human abdominal adipose tissue and cultured. The immunophenotype and adipose induced-differeneiation were identified, and the third generation cells were collected. The collected cells were assigned to 1 × 10^-8, 1 × 10^-7, 1 × 10^-6 mol/L insulin groups according to the concentration of added insulin. When cells grew into 70% confluence in conventional medium, ADSCs were cultured further in serum-free DMEM containing insulin in different concentrations for 3 days. ADSCs cultured in medium without insulin were used as control group. Secretion amount of vascular endothelial growth factor （VEGF） and hepatocyte growth factor （HGF） of ADSCs were determined by enzyme-linked immunosorbent assay. The effects of the supernatant fluid of ADSCs＇ nutrient solution on the proliferation and collagen synthesis of the cultured fibroblast were detected by MTT chromatometry and hydroxyproline chromatometry. Results The secretion amounts of VEGF and HGF of ADSCs in 1 × 10^-8 and 1 × 10^-7 mol/L insulin groups [ （471 ±41, 762 ± 66 ng/L） , （643±64, 930± 67 ng/L） , respectively] were significantly higher as compared with those in control group （286 ±47, 577 ± 84 ng/L） （ P 〈 0.05 or P 〈0.01）. No change oecurred in the secretion amount of VEGF and HGF of ADSCs in 1 × 10^-6 mol/L insulin group （ P 〉0.05）. The supernatant fluid of ADSCs＇ nutrient medium of 1 × 10 ^-8 ,1 × 10^-7 mol/L insulin groups showed obvious stimulative effect on the proliferation and collagen synthesis of fibroblasts, and it was most obvious in the 1× 10^-7 mol/L group （ P 〈0.05 orP 〈0.01）. Conclusions Insulin in the concentrations of 1 ×10^-8 and 1× 10^-7 mol/L can notably promote ADSCs＇ function of secreting VEGF and HGF.

关 键 词：胰岛素 干细胞 成纤维细胞 细胞增殖 胶原合成 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]