Flavonoids of puerarin protects vascular endothelial cells against hydrogen peroxide induced injury via up-regulating PI3K/AKT
Received:June 22, 2017  Revised:August 28, 2017
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DOI:10.11915/j.issn.1671-5403.2017.12.218
Key words:flavonoids of puerarin  endothelial cell injury  apoptosis  autophagy
Author NameAffiliationE-mail
YI Jun Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
LI Yang Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China ganfang9876@163.com 
ZHU Qing-Lei Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
YIN Tong Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
YANG Jie Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
LIU Jie Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
ZHAO Xiao-Jing Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
ZHOU Hao Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China  
CHEN Guang-Hui Department of Cardiology, Chinese PLA General Hospital, Beijing 100853, China 13910669498@163.com 
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Abstract:
      Objective To investigate the protective effect of total flavonoids in puerarin (PR) on oxidant injury in endothelial cells (ECs) induced by hydrogen peroxide (H2O2) and the possible mechanism. Methods ECs were isolated from the aorta of male Sprague-Dawley rats by adherent method. Cell viability was assessed by MTT colorimetry, and the effect of PI3K inhibitor LY294002 on the proliferation was also measured. The cell apoptosis was detected with flow cytometry. Fluorescence microscopy was used to observe the autophagic marker, microtubule-associated protein 1 light chain 3 (LC-3) in the 3 groups of cells. Western blotting was employed to detect the expression of PI3K and AKT in presence or absence of LY294002. One-way ANOVA was used to make compar-ison between groups. Results The cell viability was significantly lower in the H2O2+PR ECs with LY294002 treatment than those without[(2.07±0.08) vs (1.69±0.04), P<0.05]. Flow cytometry showed that the apoptosis rate was significantly higher in the H2O2 injured cells and the injured cells with PR treatment than the control cells (P<0.05), but PR treatment reduced the apoptosis rate in the H2O2 cells (13.26% vs 25.47%, P<0.05). Compared with the control cells, the total intracellular LC3-GFP fluorescence intensity was enhanced in the H2O2 ECs and the H2O2+PR ECs (P<0.05), with that of the latter group more significant (P<0.05). The protein levels of PI3K and AKT were up-regulated significantly in H2O2+PR ECs than the control cells (P<0.05), while the increases were blocked by LY294002 treatment (P<0.05). Conclusion PR protects ECs from H2O2 induced injury and promotes cell survival through inhibiting cell excessive apoptosis and autophagy, which may be associated with up-regulation of PI3K/AKT expression.
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