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目的观察睾酮对外周血内皮祖细胞(EPCs)分泌血管内皮生长因子(VEGF)的影响。方法取健康成年男性外周血,体外分离培养EPCs,以双荧光染色法鉴定后分为五组:A组为空白对照;B、C和D组分别加入睾酮1、10和100nmol/L;E组以磷脂酰肌醇-3-激酶/丝苏氨酸蛋白激酶(PI3K/Akt)信号通路抑制剂LY294002 40μmol/L预处理1h后,加入睾酮10nmol/L。培养48h后,ELISA法检测各组细胞上清液中VEGF蛋白量,RT-PCR检测VEGF mRNA表达。结果 B、C和D组VEGF蛋白表达量分别为(173.68±26.24)pg/ml、(252.25±9.33)pg/ml和(296.38±18.31)pg/ml,VEGF mRNA表达分别为9.19±3.48、11.05±2.19和14.87±7.36,均明显高于A组的(106.61±18.26)pg/ml和0.13±0.16(P<0.05)。E组的VEGF蛋白和mRNA表达与A组相仿(P>0.05)。结论睾酮可能通过PI3K/Akt通路促进外周血EPCs分泌VEGF。
Objective To observe the effect of testosterone on the secretion of vascular endothelial growth factor (VEGF) from peripheral blood endothelial progenitor cells (EPCs). Methods Peripheral blood was collected from healthy adult male. EPCs were isolated and cultured in vitro. The cells were identified by double-fluorescent staining and divided into five groups: group A was blank control; group B, C and D were added testosterone 1,10 and 100nmol / After pretreatment with 40μmol / L PI3K / Akt signaling pathway inhibitor LY294002 for 1h, testosterone 10nmol / L was added. After cultured for 48h, the amount of VEGF protein in the supernatant of each group was detected by ELISA, and the expression of VEGF mRNA was detected by RT-PCR. Results The protein expression of VEGF in groups B, C and D were (173.68 ± 26.24) pg / ml, (252.25 ± 9.33) pg / ml and (296.38 ± 18.31) pg / ml respectively. The mRNA expression of VEGF was 9.19 ± 3.48, ± 2.19 and 14.87 ± 7.36, respectively, which were significantly higher than those in group A (106.61 ± 18.26) pg / ml and 0.13 ± 0.16 (P <0.05). The expression of VEGF protein and mRNA in group E was similar to that in group A (P> 0.05). Conclusion Testosterone may promote the secretion of VEGF by peripheral blood EPCs through the PI3K / Akt pathway.