Protective effects and its mechanism of lipoxins on human umbilical vein endothelial cells under hypoxia
10.3760/cma.j.issn.0529-567x.2009.04.011
- VernacularTitle:脂氧素A4对缺氧损伤的人脐静脉内皮细胞的保护作用
- Author:
Songjun LIU
;
Yongsheng LI
;
Xiaoyan ZHOU
;
Lei CAI
;
Xiaoli LIU
;
Yanjun HUANG
;
Duyun YE
;
Yinping HUANG
- Publication Type:Journal Article
- Keywords:
Hypertension,pregnacy-induced;
Umbilical veins;
Endothelial cells;
Anoxia;
Lipoxins;
Calcium
- From:
Chinese Journal of Obstetrics and Gynecology
2009;44(4):281-284
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate protective effects and mechanisms of lipoxinA4(LXA4)on human umbilical vein endothelial cells(HUVEC)under hypoxia in vitro.Methods The HUVEC culture were divided into groups as followed:added M199 cudture medium as normal contml groups,added CoCl2 to mimic hypoxia in vitro as hypoxia group and added different concentrations of LXA4(1,10,100 nmol/L)were added to the induced hypoxiai HUVEC as agents intervention group.Morphological changes of HUVEC were observed by using inverted phase contrast mieroscope.The influence of LXA4 on cell survival was investigated by methyl thiazolyl tetrazolium(MTT) assaying method after the treatment with different concentrations of LXA4 and 100 nmol/L lipoxinA4 according to different time(4,8,12 and 24 hours).The expression of von-willebrand factor (vWF) was detected by immunocytoehemistry method. The changes of cytosolic Ca2+ were measured by laser scanning confocal microscope. Results ( 1 ) Morphological changes:the cells under hypoxia lost its normal shapes and showed necrosis, while the cells cocuhured with 100 nmol/L LXA4 were normal appropriately. (2)Survival rate: the survival rates of HUVEC under hypoxia was (40. 1±3.9) % and increased to ( 52. 9 ± 1.4) %, (64. 1 ± 3. 3 ) %, ( 76. 6 ± 1.6) % respectively when added with LXA4 with concentration of 1,10, 100 nmol/L into culture medium. There was significant different survival rate when compared with that of hypoxia group. (3) The level of vWF: The expression of vWF was decreased with the increasing concentrations of LXA4 added into culture medium, the gray values were 203.9 ±0. 7 in 1 nmol/L,204.6 ±0. 9 in 10 nmoL/L,191.8 ±0. 5 in 100 nmol/L respectively, which reached statistical difference in comparison with that of hypoxia groups (P<0. 05). (4) Confocai analysis:the intracellular free Ca2+ concentrations of HUVEC were intensified with LXA4 treatment. Conclusions LXA4 plays an important role in keeping the normal shape of HUVEC under hypoxia, can enhance survival of hypoxial HUVEC and decrease the level of vWF in cytoplasm. The protective mechanism might be via decreasing mitochondria Ca2+ overload and increasing cytoplasm Ca3+ by nucleus Ca2+ transference.
