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Cardiovascular disease basic research
The experimental study of protective effects of atrial natriuretic peptide (ANP) injection on the cultured human umbilical vein endothelial cells injured by hypoxia-reoxygenation
  1. Zhangqiang Chen,
  2. Chang Yao
  1. Jiangxi Provincial People's Hospital, 300 Guangzhou Road, Nanjing, Jiangsu, China

Abstract

Objective To study the protective effects of Atrial Natriuretic Peptide (ANP) Injection on the cultured human umbilical vein endothelial cells injured by hypoxia-reoxygenation.

Methods Hypoxia-reoxygenation group (n=6): cells were cultured in the hypoxia box for 6 h, then reoxygenation for 6 h; 3. hypoxia-reoxygenation+different concentrations of ANP Group: 0.001 μg/ml (n=6); 0.005 μg/ml (n=6); 0.01 μg/ml (n=6); 0.05 μg/ml (n=6); 0.1 μg/ml (n=6), and also hypoxia cultured for 1 h and reoxygenation 6 h; the cells in the cell morphology were observed under inverted microscope, cell culture supernatant obtained in each group were measured {malondialdehyde (MDA), lactate dehydrogenase (LDH), nitric oxide (NO) and endothelin (ET) content, the final results of statistical analysis conducted.

Results The dehydrogenase (LDH) activity in hypoxia-reoxygenation group was significantly higher compared with the control group (69.35±5.66 vs 31.04±3.43, p<0.01); LDH activity in drug intervention group was significantly decreased compared with hypoxic reoxygenation (p<0.01). MDA content in hypoxia-reoxygenation group was significantly higher than that in the control group (5.94±0.58 vs 1.69±0.16, p<0.01), but MDA content in drug intervention group was significantly lower than that in hypoxia-reoxygenation group (p<0.01), during the range of ANP concentration from 0.001 μg/ml to 0.05 μg/ml), MDA content in the culture medium concentration showed negative correlation with ANP concentration in the ANP group (p<0.01). Levels of intracellular SOD in hypoxia-reoxygenation group were significantly lower than that in the control group (15.74±2.17 vs 47.08±4.23, p<0.01) and was significantly higher in drug treatment group than in hypoxia-reoxygenation group (p<0.01), When ANP concentration range from 0.001 μg/ml to 0.05 μg/ml, each sub-group correlation analysis showed medium concentration of SOD content was positively correlated with ANP concentration in ANP intervention group (p<0.01); Nitric oxide (NO) levels in cell culture medium in hypoxia group was significantly lower than that in the control group (36.81±3.78 vs 89.78±7.01, p<0.01), ET-1 content was significantly higher than that in the control group (1368.64±99.07 vs 305.52±35.30 p<0.01); Correlation analysis showed that: ANP concentration ranged from 0.001 μg/ml to 0.05 μg/ml, in each sub-group of ANP intervention group, NO levels were positively correlated with ANP (p<0.01), while the concentration of ET-1 concentration was negatively correlated with ANP concentration (p<0.01).

Conclusion ANP could reduce the concentration of LDH and MDA content in the culture medium and increased human umbilical vein endothelial cell SOD content; ANP improved the content of NO and reduced the content of ET-1 after human umbilical vein endothelial cells undergoing hypoxia-reoxygenation injury, thus playing an important role in protecting the endothelial function.

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