睾酮对外周血内皮祖细胞分泌血管内皮生长因子的影响

Effect of testosterone on vascular endothelial growth factor in endothelial progenitor cells of peripheral blood

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 vascular endothelial growth factor（VEGF） in endothelial progenitor cells （EPCs） of peripheral blood. Methods EPCs obtained from peripheral blood of healthy male adults were cultured in vitro and identified by double fluorescent staining. EPCs were assigned into 5 groups. Group A was as blank control. Groups of B,C and D were treated with testosterone 1 nmol/L, 10 nmol/L and 100 nmol/L, respectively. Group E was pretreated with phosphateidylinositol 3 kinase/serine-threonine kinase（PI3K/Akt）signaling pathway inhibitor LY294002 40 μmol/L for 1 h before treated with testosterone 10 nmol/L. After cultured for 48 h, the expressions of VEGF protein and VEGF mRNA were detected by ELISA and RT-PCR. Results In groups of B,C and D,the expressions of VEGF protein were （173.68±26.24） pg/ml, （252.25±9.33） pg/ml and （296.38±18. 31） pg/ml, and the expressions of VEGF mRNA were 9.19±3. 48, 11.05±2.19 and 14.87±7. 36, respectively,which were all higher than （106.61± 18. 26） pg/ml and 0. 13±0.16 in group A（P〈0. 05）. There were no significant differences in the expressions of VEGF protein and VEGF mRNA between group E and group A（P〈0. 05）. Conclusion Testosterone may promote VEGF secretion of the EPCs through PI3K/Akt signaling pathway.