VEGF经PI3K-Akt-Bad通路降低冻存后EOCs凋亡率

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Abstract Objective: To investigate the impact of vascular endothelial growth factor (VEGF) on the apoptosis rate in cryopreservation endothelial outgrowth cells (EOCs), as well as the relationship between VEGF and the PI3K-Akt-Bad signaling pathway, to research the mechanism of VEGF reducing the apoptosis rate in cryopreservation EOCs. Methods: The mononuclear cells were harvested from umbilical cord blood, induced into EOCs and expanded in vitro. The endothelial characteristics of the EOCs were identified by immunocytochemical staining and fluorescence staining. The second generation of EOCs were devided into group W (cryoperserved after wortmannin intervention for 24 hours), group W+V (cryopreserved with 50 μg/L VEGF after wortmannin intervention for 24 hours), group BC (control group), and group V (cryopreserved with 50 μg/L VEGF). All these groups were resuscitated after being cryopreservated at -80 ℃ for 24 hours. Subsequently, apoptosis rates were detected by flow cytometry, and the expressions of p-Akt, Bad, and Caspase 3 were measured using Western-blot test. Results: The cells cultured by adherent method showed multiple endothelial characteristics. Although cryopreservation increased the apoptosis rate of EOC (5.36%±0.27%), VEGF protected cells from deep hypothermia and thus reduced the apoptosis rate of recovery cells (3.36%±0.27%, P<0.05). Inhibition of PI3K by wortmannin decreased the protection of VEGF on the EOCs and increased the cellular apoptosis rate (8.34%±0.57%, P<0.05). Western-blot test showed elevation of p-Akt expressions and decline of Bad and Caspase 3 expressions in EOCs cryopreserved with VEGF (P<0.05). Under the PI3K inhibition by wortmannin, the expression of p-Akt was down-regulated while expressions of Bad and Caspase 3 were up-regulated (P<0.05). Conclusion: VEGF decreases the apoptosis rate of cryopreserved EOCs partly via PI3K-Akt-Bad signal pathway.

Key words： vascular endothelium growth factor endothelial outgrowth cells cryopreservation apoptosis signaling pathway

Received: 10 July 2014

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	LI Zhengzheng1
	LIN Yinuo2
	GUO Yanhui2

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https://xb.wmu.edu.cn/EN/ OR https://xb.wmu.edu.cn/EN/Y2014/V44/I12/882

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