舒洛地特对低氧状态下人真皮微血管内皮细胞凋亡的影响

Effect of sulodexide on apoptosis of human dermal microvascular endothelial cells exposed to hypoxia

目的:探讨舒洛地特(sulodexide,SDX)对低氧状态下人真皮微血管内皮细胞(human dermal microvascular endothelial cells,HDMECs)凋亡的影响及其分子机制。方法:对HDMECs进行分组培养:常氧对照组在常氧状态下培养;低氧对照组于1%O_2、5%CO_2、37℃条件孵箱培养24 h;处理组用不同浓度(0.25、0.5和1 LSU/m L)舒洛地特作用于HDMECs低氧培养24 h。CCK-8法检测细胞存活率;流式细胞术检测细胞凋亡率;caspase-3活性检测试剂盒测定细胞内caspase-3活性;Western blot和real-time PCR检测促凋亡因子P53、Bax和caspase-3及凋亡抑制因子Bcl-2的蛋白及m RNA表达。结果:低氧状态下舒洛地特可提高HDMECs生长活力,降低其凋亡率,抑制促凋亡因子P53、Bax和caspase-3的表达以及caspase-3的活性,提高抑凋亡因子Bcl-2的表达。结论:舒洛地特可以抑制低氧状态下人真皮微血管内皮细胞的凋亡,其分子机制与抑制线粒体凋亡通路有关。

AIM:To investigate the effect of sulodexide(SDX)on the apoptosis of human dermal microvascular endothelial cells(HDMECs)exposed to hypoxia and its underlying mechanism.METHODS:The HDMECs were cultured and divided into normoxia control group cultured under normoxic condition;hypoxia control group cultured in a humid incubator maintained at 37℃with 5%CO2 and 1%O2 for 24 h;treatment groups treated with SDX at 0.25,0.5 and 1 LSU/mL for 24 h under hypoxic condition.The cell viability was measured by CCK-8 assay.The apoptotic rate of the HDMECs was analyzed by flow cytometry.The activity of caspase-3 in HDMECs was examined by caspase-3 activity assay kit.The expression of pro-apoptotic factor P53,caspase-3,Bax and anti-apoptotic factor Bcl-2 at mRNA and protein levels was determined by real-time PCR and Western blot.RESULTS:SDX increased the viability of HDMECs exposed to hypoxia,but also decreased the apoptosis.Furthermore,SDX down-regulated the expression of pro-apoptotic factor P53,Bax and caspase-3 at mRNA and protein levels as well as the activity of caspase-3,while the expression of anti-apoptotic factor Bcl-2 was up-regulated.CONCLUSION:SDX significantly increases the viability and decreases the apoptosis of HDMECs exposed to hypoxia.Inhibition of the mitochondrial apoptosis pathway may be involved in the underlying mechanism.