摘要
在牛主动脉内皮细胞培养的基础上建立缺氧、再灌注模型,观察内皮细胞谷胱甘肽(GSH)、丙二醛(MDA)和上清液中一氧化氮(NO)、乳酸脱氢酶(LDH)含量的变化,同时应用电镜技术观察细胞形态的改变。实验发现缺氧2小时(100％N_2)、再灌注30分钟(95％O_2、5％CO_2)血管内皮细胞内GSH含量显著下降,MDA含量显著上升。透射电镜下见内皮细胞内空泡增多,细胞膜也有不同程度损害。在缺氧过程中应用东莨菪碱(scopolamine)可预防内皮细胞GSH消耗,减少MDA生成,并减少上清液NO、LDH释放,均具有统计学意义。我们认为scopolamine这种作用机制与其抗脂质过氧化作用有关。scopolamine还通过抗脂质过氧化作用,减少脂质过氧化物形成,减少GSH消耗。scopolamine减少NO释放的机制,可能与防止再灌注过程中细胞钙过载有关。
This study was designed to determine the protective effect of scopolamine and catalase on the arterial endothelial cells during reoxygenation. The culture of endothelial cells from bovine aorta subjected to ischemia for 2 hours and reoxygenation for 30 minutes was used as a model for inves-tigaion. In the experimental group, scopolamine was added to the arterial endothelial cells culture medium, Hanks solution containing no sugar or oxygen; or in the control group no medicine was added to the ordinary Hanks solution. The alteration of glutathione ( GSH ) and malondialdehyde (MDA) of endothelial cells, and nitric oxide (NO) and lac-tate dehydrogenase (LDH) in the supernatant fluid were determined. The results demonstrated the following changes: (1) GSH in the control group significant reduction from the preischemia value 44. 5 + 2.45 to postischemia and reoxy genation value 35.08 + 5.72ug/mg protein, vs the post-ischemia and reoxygenatior value of 46. 67+3.20ug/ mg protein (scopolamine) and 43.38+2.98ug/mg protein (catalase). The differences between the GSH changes of control group and medicine treated group are statistically significant (P<0. 05). (2)MDA: in the control group, the MDA contents increased from preischemic value 1. 13+0.12nM/ 106 cells to 1.84+0.06nM/106 cells after ischemia and reoxygenation (P<0.01) vs 1. 19+0.07nM/106 cells (with scopolamine 10-5mol/L) and 1. 44+0.007nM/106cells (with scopolamine 10-7mol/L) after ischemia and reoxygenation. As the last two values compared with the corresponding value of the control group, P value was<0. 01 and <0. 05 respectively. Addition of catalase (1200u/ml) to the culture medium could aiso markedly reduce the production of MDA as compared with that of the control group (P<0.05); (3) NO: in the control group, the NO content in the supernatant fluid of preischemia arterial endothelial cells was 46. 96 ?22.17ng/ml which rose to 112.24+43.6ng/ml (P<0.05) after subiected ischemia and reoxygenation. Addition of scopolamine 10-5mol/L or 10-7mol/L could obviously decrease the NO content to 41. 31+19. 55ng/ml and 46. 96?22. 17ng/ml respectively (P values were all<0. 05). (4) LDH: in the control group the LDH content in the supernatant fluid showed extremely high release, from preischemia value 77. 65+10. 4U/ml to 404. 34+37. 88U/ml(P <0.001). In the group with scopolamine 10-5mol/L or 10-7 mol/L, the release of LDH was very markedly reduuced to 128.36+13.86U/ml and 226. 49+16.23U/ml, P values were all<0.01 as compared with the corresponding value of the control group. Addition catalase (1200U/ml) could also decrease the release of LDH to 287. 5 + 17. 76U/ml (P< 0.05).
The morphological changes of endothelial cells were studied under transmission electron microscope. Ultrastruc-tural changes including cytoplasmic vacuolization and various degrees of cellular membrane injures were observed and in a lesser degree in the experimental group.
The afore mentional date indicate the protective effects of scopolamine against ischemia and re-oxygenation injures in the cultured arterial endotheliol cells are obvious.
出处
《中华胸心血管外科杂志》
CSCD
北大核心
1995年第5期306-308,共3页
Chinese Journal of Thoracic and Cardiovascular Surgery
关键词
内皮细胞
缺氧
再灌注
东莨菪碱
动脉
保护作用
Arterial endothelial cell culture
Anoxia
Re-oxygenation
Glutathione
Nitric oxide
