C-反应蛋白对脐静脉内皮细胞凋亡和P选择素表达的影响

The Effect of C-reactive Protein (CRP) on Human Umbilical Vein Endothelial Cells (HUVECs) Apoptosis and the Expression of P-selectin

目的:观察C-反应蛋白(C-reactive protein,CRP)对人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVECs)凋亡和P选择素(P-selectin)表达的影响。方法:将HUVECs与不同浓度CRP(0ug/ml、5ug/ml、25ug/ml、50ug/ml、100ug/ml)在培养基中孵育24h后为CRP剂量效应组;0ug/ml、50ug/mlCRP作用3h、6h、12h、24h为时间效应组;分别收集细胞和上清液。采用流式细胞仪检测细胞凋亡率,用酶联免疫吸附试验检测上清液中的P选择素含量。结果:(1)与对照组(0ug/mlCRP)相比,5ug/ml的CRP培养内皮细胞24h细胞凋亡率显著增加(12.16±3.69%,P<0.01),50ug/ml时细胞凋亡率到达顶峰(27.91±3.11%,P<0.01);50ug/mlCRP孵育不同时间,细胞凋亡率从3h明显增加(2.71±0.93%,P<0.01),24h时细胞凋亡率达顶峰(27.91±3.11%,P<0.01)。(2)与对照组(0ug/mlCRP)相比,5ug/ml的CRP即可显著增加内皮细胞P选择素表达(15.93±3.77ng/ml,P<0.01),100ug/ml时P选择素表达到达顶峰(86.35±7.35ng/ml,P<0.01);50ug/mlCRP孵育不同时间,P选择素表达从3h明显增加(34.33±5.01ng/ml,P<0.01),6h时P选择素表达达顶峰(79.28±7.63ng/ml,P<0.01)。结论:CRP以剂量及时间依赖的方式促进内皮细胞凋亡及P选择素的表达。

Objective To observe the effect of ClIP on HUVECs apeptosis and the expression of P-seleetin. Methods Through incubation of HUVECs with different concentration of CRP（0ug/ml,5ug/ml,25ug/ml,50ug/ml, 100ug/ml） for 24hours, to observe the dose effect of CRP on HUVECs apoptosis. Through incubation of HUVECs with ClIP （50ug/ml） for different time （3 hours,6 hours, 12 hours,24 hours）, to observe the time effect of CRP on HUVECs apoptosis. Cells apeptosis analyzed by flow cytometry and P-seleetin measured by Enzyme-linked immunosorbent assay （ELISA）. Results （1） Compared with control group（0ug/ml CLIP）, ClIP caused a significant increase in the rate of HUVECs apoptosis at a dose as low as 5ug/ml（12.16% ± 3.69%, P 〈 0.01） and it reached to the peak value at 50ug/ml （27.91 % ±3.11%, P 〈 0.01 ）. The apoptosis rate was markedly elevated as early as 3 hours （2.71% ±0.93%, P 〈 0.01） after HUVECs incubation with ClIP 50ug/ml and it reached to the peak value at 24hours （27.91 ± 3.11, P 〈 0.01 ）. （2）. Compared with control group（0ug/nd CLIP）, ClIP caused a significant increase in the expression of P-seleetin at a dose as low as 5ug/ml（ 15.93ng/ml ＋ 3.77ng/ml, P 〈 0.01 ） and it reached to the peak value at CRP 100ng/ml （ 86.35ng/ml ±7.35ng/ml, P 〈 0.01）. After incubation with CRP 50ug/ml, the expression of P-seleefin was elevated as early as 3 hours （34.33ng/ml ± 5.01ng/ml, P 〈 0.01） and it reached to the peak value at 6hotrrs （79.28ng/ml ±7.63ng/ml, P 〈 0.01 ）. Conclusions CRP induced HUVECs apeptosis and upregulated the expression of P-seleetin in a dose-and time-dependent manner.