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目的:体外观察100 ng/m L血管紧张素-Ⅱ(Angiotensin-Ⅱ,Ang-Ⅱ)处理后的人脐带间充质干细胞(human umbilical cord MSCs,h UCMSCs)上清液对损伤的人脐静脉内皮细胞(human umbilical vein endothelia cells,HUVEC)增殖、凋亡的影响。方法:CCK8法检测HUVEC增殖情况;倒置显微镜下观察HUVEC形态;吖啶橙/溴化乙锭染色法(acridineorange/ethidium bromide,AO-EB)、Hoechst检测HUVEC凋亡情况。结果:CCK8实验结果显示,加入100 ng/m L Ang-Ⅱ处理后的h UCMSCs上清液组HUVEC增殖速度在各时间点显著快于其他两组。倒置显微镜下观察细胞的形态:加入100 ng/m L Ang-Ⅱ处理后的h UCMSCs上清液组HUVEC的形态最佳且凋亡数最少。用各组细胞上清液对HUVEC培养24 h、48 h、72 h后,AO-EB、Hoechst染色结果显示:加入100 ng/m L Ang-Ⅱ处理后的h UCMSCs上清液组,在各个时间点HUVEC出现凋亡细胞或死亡细胞的数最少。结论:100ng/m L Ang-Ⅱ预处理后的h UCMSCs上清液可以促进HUVEC增殖、抑制HUVEC凋亡。
OBJECTIVE: To observe the effects of human umbilical cord MSCs (h UCMSCs) supernatants treated with 100 ng / m L Angiotensin-Ⅱ (Ang-Ⅱ) on human umbilical vein injury (UUV) on the proliferation and apoptosis of human umbilical vein endothelia cells (HUVECs). Methods: The proliferation of HUVECs was detected by CCK8 method. The morphology of HUVECs was observed under inverted microscope. The apoptosis of HUVECs was detected by acridine orange / ethidium bromide (AO-EB) and Hoechst staining. Results: The results of CCK8 assay showed that the proliferation rate of HUVECs in h UCMSCs supernatant treated with 100 ng / m L Ang-Ⅱ was significantly faster than the other two groups at each time point. The morphology of the cells was observed under an inverted microscope. HUVECs in the supernatant of h UCMSCs treated with 100 ng / m L Ang-Ⅱ had the best morphology and the lowest number of apoptotic cells. The results of AO-EB and Hoechst staining showed that after treated with 100 ng / m L Ang-Ⅱ for 24 hours, 48 hours and 72 hours, the supernatant of h UCMSCs cultured in each group At the time point, the number of apoptotic cells or dead cells in HUVEC was the least. Conclusion: The supernatant of h UCMSCs pretreated with 100ng / m L Ang-Ⅱ can promote the proliferation of HUVEC and inhibit the apoptosis of HUVEC.