【摘要】 目的 研究人尿源干细胞(USCs)对糖基化终末产物(AGEs)损伤的海绵体血管内皮细胞(CCECs)的保护作用。方法 流式细胞仪鉴定大鼠阴茎海绵体血管内皮细胞表面标志物；收集3名健康成年男性无菌尿液9份，分离、原代培养USCs,流式细胞仪鉴定USCs细胞表面标志物。将实验分为六组，分别为正常对照组、干细胞上清组、干细胞组、AGEs组、AGEs+干细胞上清组、AGEs+干细胞组。在相应组别加入AGEs(200μg/mL)处理24h后，Annexin V法检测各组细胞凋亡情况；Western Blot检测Bcl2-Associated X的蛋白质(Bax)、B淋巴细胞瘤-2基因(Bcl2)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-PX)、活性氧(ROS)、内皮型一氧化氮合酶(eNOS)蛋白含量表达水平。结果 成功分离培养人USCs,鉴定CD24、CD44、CD73、CD90呈强阳性，CD105弱阳性，CD31、CD34、CD45呈阴性。流式细胞仪成功鉴定CCECs表型CD31阳性、CD90阴性。AGEs组凋亡情况高于正常对照组，AGEs+干细胞组及AGEs+干细胞上清组凋亡情况低于AGEs组，差异具有统计学意义(P<0.05)。AGEs组Bcl2、SOD、GSH-PX和eNOS蛋白含量表达均低于正常对照组；AGEs+干细胞组及AGEs+干细胞上清组Bcl2、SOD、GSH-PX和eNOS蛋白含量表达强于AGEs组；AGEs组Bax、ROS蛋白含量表达高于正常对照组，AGEs+干细胞组及AGEs+干细胞上清组Bax、ROS蛋白含量表达低于AGEs组(P<0.05)。结论 AGEs使海绵体血管内皮细胞的凋亡增加，USCs能改善相关指标，对受AGEs损伤的CCECs具有保护作用。更多还原

【Abstract】 Objective To investigate the protective effect of human urine-derived stem cells(USCs) on advanced glycation end products(AGEs) damaged cavernous vascular endothelial cells(CCECs). Methods The surface markers of vascular endothelial cells in the cavernosum of the penis of rats were identified by flow cytometry. 9 samples of sterile urine were collected from 3 healthy adult males, USCs were isolated and cultured in primary culture, and USCs cell surface markers were identified by flow cytometry. The experiment was divided into six groups: normal control group, stem cell supernatant group, stem cell group, AGEs group, AGEs+ stem cell supernatant group, AGEs+ stem cell group. Apoptosis was detected by Annexin V assay after the corresponding groups were treated with glycation end products(200 g/mL) for 24 h. Western Blot was used to detect the protein expression levels of Bcl2-associated X(Bax), B-cell lymphoma-2(Bcl2), superoxide dismutase(SOD), glutathione peroxidase(GSH-PX), reactive oxygen species(ROS) and endothelial nitric oxide synthase(eNOS). Results Human USCs were isolated and cultured successfully. CD24, CD44, CD73 and CD90 were strongly positive, CD105 was weakly positive, and CD31, CD34 and CD45 were negative. CD31 positive and CD90 negative CCECs phenotypes were successfully identified by flow cytometry. Apoptosis in AGEs group was higher than that in normal control group, and that in AGEs+stem cell group and AGEs+stem cell supernatant group was lower than that in AGEs group(P<0.05).The protein expression levels of Bcl2, SOD, GSH-PX and eNOS in AGEs group were lower than those in normal control group, and that in AGEs+stem cell group and AGEs+stem cell supernatant group were lower than those in AGEs group; the protein expression levels of Bax and ROS were higher than those in normal control group, while the protein expression levels of Bax and ROS in AGEs+stem cell group and AGEs+stem cell supernatant group were lower than those in AGEs group(P<0.05). Conclusions AGEs can increase the apoptosis of spongiform vascular endothelial cells. USCs can improve the related indexes, and has a protective effect on AGEs damaged CCECs.更多还原