腺病毒介导的Vasostatin基因对血管内皮细胞的作用

Effect of Vasostatin Gene Mediated by Adenovirus on the Proliferation and Apoptosis of Vascular Endothelial Cells

目的研究腺病毒介导的Vasostatin基因对体外培养的血管内皮细胞增殖及凋亡的作用。方法采用MTT法观察Vasostatin基因对人脐静脉血管内皮细胞ECV304的体外细胞增殖的影响。采用透射电镜、TUNEL与Hoechst33258双重荧光染色、流式细胞仪AnnexinV/PI双染法,检测腺病毒介导的Vasostatin基因作用后人脐静脉血管内皮细胞的凋亡。结果Ad-Vasostatin(MO I分别为25和50)作用72 h后,ECV304细胞数显著少于PBS组和Ad-lacZ组(P<0.05);透射电镜下及TUNEL/Hoechst33258双重荧光染色,ECV304细胞出现典型的凋亡形态学改变。以MO I为50的Ad-Vasostatin处理ECV304细胞72 h后,细胞凋亡率为(15.70±0.84)%,PBS组为(2.54±0.83)%,Ad-lacZ组为(2.34±0.79)%,三组比较均有显著性差异(P<0.01)。结论腺病毒介导的Vasostatin基因可显著抑制人脐静脉血管内皮细胞ECV304的体外细胞增殖,同时对人脐静脉血管内皮细胞的凋亡具有诱导作用。

Objective To investigate the effect of vasostatin mediated by adenovirus on the proliferation and apoptosis of vascular endothelial cells in vitro. Methods The effects of vasostatin gene on the proliferation of ECV304 cells were measured by MTT assay. Morphological changes of ECV304 cells were observed under electron microscopy. Apoptosis was determined by Hoechst33258/TUNEL staining, and by FCM with Annexin V/P1 costaining. Results In the cellular proliferarion assay in vitro, the number of ECV304 cells in Ad-Vasostatin group was much smaller than that in Ad-lacZ group and PBS group （P 〈 0.05）. After 72 h of treatment with Ad-Vasostatin, the ECV304 cells had characteristics of apoptosis, as revealed by electron microscopy and Hoechst33258/TUNEL costaining. The apoptosis rate in Ad-Vasostatin group （15.70 ± 0.84）% was much higher than that in Ad-lacZ group （2.34±0.79）% and PBS group （2.54±0.83）% （P〈0.01）. Conclusion The vasostatin gene mediated by adenovirus has an inhibitory effect on the proliferation of human umbilical vein endothelium cells ECV304 in vitro, and could induce the apoptosis of human umbilical vein endothelium cells in vitro.