Glycine prevents apoptosis of rat sinusoidal endothelial cells caused by deprivation of vascular endothelial growth factor

被引:43
|
作者
Zhang, YJ [1 ]
Ikejima, K [1 ]
Honda, H [1 ]
Kitamura, T [1 ]
Takei, Y [1 ]
Sato, N [1 ]
机构
[1] Juntendo Univ, Sch Med, Dept Gastroenterol, Bunkyo Ku, Tokyo 1138421, Japan
关键词
D O I
10.1053/jhep.2000.16605
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
Apoptosis of sinusoidal endothelial cells (SECs) is one of the initial events in the development of ischemia-reperfusion injury of the liver. Glycine has been shown to diminish. ischemia-reperfusion injury in the liver and improve graft. survival in the rat liver transplantation model. Here, we investigated the effect of glycine on apoptosis of primary cultured rat SECs induced by vascular endothelial growth factor (VEGF) deprivation. Isolated rat SECs were cultured in EBM-2 medium supplemented with 10% fetal bovine serum (FBS) and growth factors including 20 ng/mL VEGF for 3 days. SECs at 3 days of culture showed spindle-like shapes; however, cells started shrinking and detaching from dishes by VEGF deprivation. Apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated d-uridine triphosphate (dUTP)-biotin nick end labeling (TUNEL) staining in these conditions. Control SECs contained only a few percent of TUNEL-positive cells; however, they started increasing 4 hours after VEGF deprivation, and the percentage of TUNEL-positive cells reached about 50% at 8 hours and almost 100% at 16 hours after VEGF deprivation. Interestingly, this increase in TUNEL-positive cells after VEGF deprivation was prevented significantly when glycine (1-10 mmol/L) was added to the medium, the levels being around 60% of VEGF deprivation without glycine, Furthermore, strychnine (1 mu mol/L), a glycine receptor antagonist, inhibited this effect of glycine, suggesting the possible involvement of the glycine receptor/chloride channel in the mechanism. Moreover, Bcl-2 protein levels in SECs were decreased 8 hours after VEGF deprivation, which was prevented almost completely by glycine, It is concluded that glycine prevents apoptosis of primary cultured SECs under VEGF deprivation.
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页码:542 / 546
页数:5
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