人尿源干细胞对糖基化终末产物损伤的海绵体血管内皮细胞的保护作用的研究  

The protective effect of human urine-derived stem cells on cavernosum vascular endothelial cells damaged by advanced glycation end products

作　　者：赵超 刘豪圣 刘天琦 张振辉 王倩倩 李勇 彭天明 陈育纯 蒲小勇 刘久敏 ZHAO Chao;LIU Haosheng;LIU Tianqi;ZHANG Zhenhui;WANG Qianqian;LI Yong;PENG Tianming;CHEN Yuchun;PU Xiaoyong;LIU Jiumin(Master Degree Candidate of Clinical Medicine Enrolled in 2019,School of Medicine,South China University of Technology,Guangzhou 510006,Guandong,China;Department of Urology,Guangdong Provincial People′s Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,Guangdong,China;School of Medicine,South China University of Technology,Guangzhou 510006,Guandong,China)

机构地区：[1]华南理工大学医学院,广州510006 [2]广东省人民医院(广东省医学科学院)泌尿外科,广州510080

出　　处：《中国性科学》2022年第12期30-34,共5页Chinese Journal of Human Sexuality

基　　金：广东省自然科学基金(2019A1515012019)。

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。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(200g/mL)for 24h.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

关 键 词：尿源干细胞 糖基化终末产物 海绵体血管内皮细胞 

分 类 号：R697[医药卫生—泌尿科学]