目的：观察特异性5型磷酸二酯酶抑制剂（PDE5）伐地那非对人羊膜间充质干细胞（hAMSCs）生物学特性的影响。方法：分离培养hAMSCs，以终浓度为10μmol/L伐地那非预处理hAMSCs，观察伐地那非处理hAMSCs组（Vard-hAMSCs）生长形态，采用流式细胞术分析比较hAMSCs细胞免疫表型、增殖能力及抗氧化损伤能力；免疫双荧光染色计数阳性表达细胞数，计算hAMSCs向神经细胞诱导分化百分率。结果：（1）流式细胞术结果显示，hAMSCs、Vard-hAMSCs均阳性表达CD90、CD105、CD73，不表达CD45、CD34、CD19、CD11b及HLA-DR；Vard-hAMSCs组细胞S期细胞比率及增殖指数为（0．57±0．40）％和（2．20±1．60）％，与hAMSCs组[分别是（0．62±1．33）％，（1．90±1．40）％]比较，无统计学差异（P ＞0．05）；（2）Annexin V/PI双染色结果显示，经250μmol/L H2O2作用4 h后，Vard-hAMSCs组细胞凋亡率为（7．67±0．82）％，明显低于hAMSCs组（18．72±2．92）％和特异性Vardenafil阻断剂组（15．90±1．69）％（均P ＜0．05）；（3）免疫双荧光染色结果显示，在相同诱导条件下，Vard-hAMSCs组细胞MAP-2及GFAP阳性细胞百分率分别是（49．8±6．42）％和（55．2±6．10）％，均高于hAMSCs组[分别为（29±3．94）％，（32．2±3．03）％，P＜0．05]。结论：伐地那非处理一定时间可明显提高hAMSCs抗氧化损伤效应和hAMSCs向神经细胞分化的能力，但短时间内不明显影响hAMSCs生长形态、增殖能力和免疫表型。提示伐地那非处理可能作为hAMSCs移植治疗神经系统疾病的优选预处理方案。%Objective To investigate the biological properties of human amniotic mesenchymal stem cells (hAMSCs) which were preconditioned with phosphodiesterase-5 inhibitor (Vardenfil). Methods hAMSCs were in vitro isolated and cultured, hAMSCs were pre-treated with vardenfil in final concentration of 10 μmol/L. The morphology of Vard-hAMSCs was observed, and the immunological characteristics, proliferative capacity, and ability of anti-oxidative damage of hAMSCs and Vard-hAMSCs were analyzed by flow cytometry. Double labeling immunofluorescent staining was used to count the differences of differential potential between neural cells of hAMSCs and Vard-hAMSCs. Results (1)Flow cytometry revealed that both hAMSCs and Vard-hAMSCs positively expressed CD90、CD105 and CD73, and negatively expressed CD34、CD45、CD11b and HLA-DR. The SPF and PI in Vard-hAMSCs group were (0.57 ± 0.40)% and (2.20 ± 1.60)% respectively, there was no statistical significance compared with hAMSCs group; (2)After 4 hours treated by H2O2, the apoptosis rate in Vard-hAMSCs group were (7.67 ± 0.82)%,which were markedly lower than that in the hAMSCs group and specific blocker group; (3)Under the same induction condition, positive rates of MAP-2 and GFAP in Vard-hAMSCs group were (49.8 ± 6.42)%and (55.2 ± 6.10)% respectively detected by double labeling immunofluorescent staining, which were significantly higher than the control group. Conclusion The strategy that hAMSCs are treated with vandenfil can enrich the ability of anti-oxidative damage and the differential potential for neural cells in a certain time, and the morphology, immunological characteristics, proliferative capacity of Vard-hAMSCs have no significant change. It suggests that pre-treatment with vandenfil may provide a optimized experimental strategey for hAMSCs which were used to treat nervous system disease.
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