Protective Effects of Sasanquasaponin on Injury of Endothelial Cells Induced by Anoxia and Reoxygenation in vitro

Abstract

Abstract:The protective effects of sasanquasaponin, an effective compound from Chinese traditional herbs, on ischaemia and reperfusion injury in mouse hearts have been suggested through modulation of intracellular Cl -homeostasis. The effects of sasanquasaponin on injury of endothelial cells, however, induced by anoxia and reoxygenation remain unknown. Therefore, the present study attempted to observe the effects of sasanquasaponin on anoxia and reoxygenation injury in endothelial cells and investigate its putative mechanisms. Human umbilical vein endothelial cells (HUVECs) were exposed to normoxia or anoxia and reoxygenation in the absence or presence of sasanquasaponin (10.0, 1.0 and 0.1 μ mol / l). Lactate dehydrogenase activity was determined in cultured HUVECs supernatant, and malondialdehyde content, superoxide dismutase and glutathione peroxidase activities were measured in HUVECs by a colorimetric method. Neutrophil adhesion to HUVECs was assayed colorimetrically. The levels of intercellular adhesion molecule-1 and tumour necrosis factor-α were detected. The activity of nuclear factor kappa B was determined by flow cytometry. The results show that sasanquasaponin decreased the lactate dehydrogenase activity and malondialdehyde contents, and inhibited the neutrophil adhesion to HUVECs; sasanquasaponin, moreover, inhibited nuclear factor kappa B transnuclear activity, lowered tumour necrosis factor-α and intercellular adhesion molecule-1 expression levels. On the other hand, sasanquasaponin increased the mitochondrial superoxide dismutase and glutathione peroxidase activities. It is suggested that sasanquasaponin could protect HUVECs against anoxia and reoxygenation injury, and the protective mechanisms appear to be related to anti-lipoperoxidation and anti-adhesion.Ischaemia and reperfusion are likely to play an important role in the pathogenesis of a wide variety of clinical conditions such as peripheral vascular disease, stroke and myocardial infarction [1,2]. Adhesion of leucocytes to endothelial cells is involved in the progression of these diseases. There is much evidence that neutrophil adhesion is a critical factor in ischaemia and reperfusion injury, and vascular endothelium is a crucial site of ischaemia and reperfusion injury [3,4]. Data have shown that endothelial cells, under ischaemic or hypoxic conditions, present dysfunction of energy metabolism, overload of intracellular Ca 2+ concentration ([Ca 2+

Antisense Oligodeoxynucleotide Against Tissue Factor Inhibits Human Umbilical Vein Endothelial Cells Injury Induced by Anoxia-Reoxygenation

“…In addition, HUVECs viability declined and LDH activity increased to a certain degree in A/R groups, which indicated HUVECs were injured by anoxia-reoxygenation. This suggested the experimental model was reliable and successful [2].…”

“…Then the veins were washed with PBS, the collagenase solution and the wash were collected and centrifuged for 10 min at 200×g, cells were resuspended in RPMI 1640 complete medium (GIBCO) supplemented with 20% fetal bovine serum (FBS, HyClone), penicillin (100 U/ml) and streptomycin (100 μg/ml), and then were seeded on glass slides ( 1.8×10 6 cells per slide) coated with gelatin and incubated at 37°C in the presence of humidified 95% air and 5% CO 2 . Upon reaching 75% confluence, cells were subcultured by the ratio of 1:3.…”

“…50 μl of the suspension was added to 0.95 ml 0.4% trypan blue solution. The total number of living and dead cells was counted with a haemacytometer [2].…”

“…Anoxia-reoxygenation or ischemiareperfusion is a pathological process involving oxidative stress. It plays an important role in the pathogenesis of a wide variety of clinical conditions such as peripheral vascular disease, stroke and myocardial infarction, etc [1,2]. As a crucial site, endothelial cells undergo cellular activation even death following anoxia-reoxygenation or ischemia-reperfusion injury.…”

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“…Control effect of saponin against insects, pests and mites has been reported (Kawai et al, 1999). In addition, It is used as a medicine for the treatment of intestinal disorders (Huang et al, 2007) and burn injuries (Huang et al, 2005). Saponin has also been reported to exert many pharmacological effects, including antihyperlipidemic (Yoshikawa et al, 2005), antiallergic (Matsuda et al, 2010), and cardioprotective effects (Liao et al, 2009).…”