Autophagy is involved in the dysfunction of vascular endothelial induced by arsenic / 中华地方病学杂志

ABSTRACT

Objective:To explore whether autophagy is involved in dysfunction of vascular endothelial induced by sodium arsenite (NaAsO 2). Methods:Human primary umbilical vein endothelial cells were isolated and cultured. The cells were treated with different levels of NaAsO 2 [0 (control)), 2, 5, 10, 20, 30, 50 μmol/L] for 24 h, and cell viability was determined using CCK8. According to the results of CCK8, the levels of arsenite used in subsequent experiments were determined, intracellular nitric oxide (NO) content (incubated by NaAsO 2 for 4 h) was detected by flow cytometry, LC3 levels (incubated by NaAsO 2 for 0, 6, 12 and 24 h) was detected using Western blotting, and autophagosome (incubated by NaAsO 2 for 12 h) was observed by electron microscope. At the same time, human primary umbilical vein endothelial cells were pretreated with 0.1 mmol/L 3-MA (autophagy inhibitor) for 2 h, and induced by 30 μmol/L NaAsO 2, and the above detection indicators were compared with those of the 30 μmol/L NaAsO 2 group. Results:Human primary umbilical vein endothelial cells were successfully isolated and cultured. Compared with the control group [cell viability (99.97 ± 5.33)%, NO content 42 048.34 ± 789.61], the cell viability [(73.00 ± 0.86)%] and NO content (23 353.97 ± 971.85) of 30 μmol/L NaAsO 2 group were remarkably lower, and the differences were statistically significant ( P < 0.01). Incubated with 30 μmol/L NaAsO 2 at different time points 6, 12, 24 h, LC3Ⅱ levels (5.782 ± 2.789, 9.692 ± 2.222, 5.573 ± 2.941) were significantly elevated than those of control group (1.000 ± 0.383, P < 0.05), and the LC3 Ⅱ level was the highest at 12 h. After treatment with 30 μmol/L NaAsO 2 for 12 h, the number of autophagosome in cells observed under electron microscope was significantly higher than that of the control group. Compared with 30 μmol/L NaAsO 2 group [cell viability (68.78 ± 1.55)%, LC3 Ⅱ level 5.680 ± 0.545, NO content 13 025.78 ± 962.61], cell viability [ (79.54 ± 4.99) %] in 3-MA+ NaAsO 2 group was increased, the LC3Ⅱ level (3.956 ± 0.398) was decreased, and the differences were statistically significant ( P < 0.05); intracellular NO content (13 988.51 ± 1 671.07) increased, whereas the difference was not statistically significant ( P > 0.05). Conclusion:Autophagy is involved in the vascular endothelial dysfunction induced by arsenic.