Medicinal Plant

2013, v.4(04) 35-38+40

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Research on the Hypoxia Resistant Ability of Vascular Endothelial Cell Improved by Tongxinluo via the PI-3K/Akt/Hif Pathway
Research on the Hypoxia Resistant Ability of Vascular Endothelial Cell Improved by Tongxinluo via the PI-3K/Akt/Hif Pathway

Junqing LIANG;Haibo XU;Zhijun XING;Ziqian ZHANG;Bing YAO;Beipei YIN;Ziming LU;

摘要(Abstract):

[Objective]To investigate the influence of PI-3K/Akt/Hif pathway on anti-hypoxia activity of Tongxinluo by studying the effect of Tongxinluo on hypoxia induced factor in vascular endothelial cell and its upstream signal transduction pathway,the PI-3K/Akt pathway,under the hypoxia condition.[Method]Human umbilical vein endothelial cells (HUVECs) were divided into the following treatment groups: control group,Tongxinluo group,hypoxia group,and hypoxia + Tongxinluo(100 μg/mL) group. The CCK-8 kits were used to detect viability and proliferation rate of cells in each group. The protein levels of Hif,Bcl 2 ,Mcl,Bax and phosphorylated Akt were studied by immunoblotting analysis. The HUVEC cells were transiently transfected with the dominant negative mutant of Hif-1α(DN-Hif) and apoptosis rates were analyzed by flow cytometry(FCM). The HUVEC cells were transiently transfected with the dominant negative mutant of PI-3K (ΔP85) or Akt (DN-Akt) to interrogate the role of PI-3K/Akt signaling pathway in the anti-hypoxia ability of Tongxinluo on endothelial cells. [Result]Though the proliferation rate increased significantly in Tongxinluo group under the hypoxia condition compared with hypoxia group,the degree weakened notably compared with the control group. The protein levels of Hif-1α,Bcl 2 ,Mcl and phosphorylated Akt were up-regulated by Tongxinluo. Meanwhile,the expression of Bax was down-regulated. Inhibition of Hif-1α activation by DN-Hif and inhibition of the PI-3k/Akt pathway by ΔP85 or DN-Akt attenuated the rise of Hif expression and HUVEC viability induced by Tongxinluo. But still,the percentage of apoptosis of HUVEC was down-regulated to a certain extent by Tongxinluo.[Conclusion] Tongxinluo could up-regulated the protein level of Hif,promote the expression of apoptosis related factors,improve the cell viability and eventually reduce the apoptosis rate,which may be one of the molecular mechanisms that Tongxinluo improved the anti-hypoxia ability of vascular endothelial cells. And futher study showed that Tongxinluo activated the PI-3K/Akt-dependent signaling pathway to up-regulate the expression of Hif.

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基金项目(Foundation): Supported by the National Fundamental Research and Development Plan(National 973 Project) (No.2012CB518606 and No.2005CB523300);; the Youth Fund Program of National Natural Science Foundation (No.81102768);; the Subject for Developing New Drugs of Major Science Project in the "Twelfth-Five Year Plan" (No.2011ZX 0940-020)

作者(Author): Junqing LIANG;Haibo XU;Zhijun XING;Ziqian ZHANG;Bing YAO;Beipei YIN;Ziming LU;

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