摘要目的 观察ARA290对人脐静脉内皮细胞(HUVEC)低温缺氧复氧损伤中的保护作用.方法 建立HUVEC细胞低温缺氧复氧模型;原代培养72 h后,并随机分为4组:(1)对照组(Control):更换不含血清的1640培养液继续4℃培养10h;(2)缺氧/复氧组(H/R):更换不含血清的1640培养液4℃缺氧保存6h/复氧4 h;(3) ARA290组:更换经含10 nmol/ml ARA290的UW器官保存液4℃保存6h/复氧4 h;(4)UW器官保存液组(UW):更换UW器官保存液并4℃缺氧保存6h/复氧4h;采用噻唑蓝(MTT)比色法检测细胞活力、膜联蛋白V(Annexin V)/异硫氰酸荧光素(FITC)、碘化丙锭(Pl)双染色流式细胞仪检测细胞凋亡.结果 (1)经过ARA290预处理的细胞存活率(89.5±7.1)％,显著高于UW组(76.3±6.0)％和H/R组(71.4±5.9)％(P＜0.01),而略低于对照组的(97.3±6.8)％(P＜0.05);(2)细胞凋亡率H/R组(13.13±0.94)％、对照组(1.15±0.18)％、ARA290组(5.34±0.61)％、UW组的(11.45±0.71)％,ARA290预处理组的细胞凋亡率,与其余3组比较差异有统计学意义(P＜0.05).结论 促红素衍生肽ARA290可对HUVEC细胞的低温缺氧复氧损伤有保护作用,可抑制其凋亡.

abstractsObjective To investigate the protective effect of ARA290,a non-erythropoietic erythropoietin (EPO) derivative,on the binding activity of nuclear factor-κB (NF-κB) and the expression of adhesion molecular and chemokines of human umbilicalvein endothelial cells (HUVEC) induced by hypoxia/reoxygenation (H/R) injury.Methods The HUVEC H/R injury model was built.The HUVEC monolayers were randomly divided into 4 groups:control,H/R,ARA290,UW.All groups,except control group,were subjected to hypoxia for 6 h in UW or UW containing 10 nmol/ml ARA290 at 4℃ followed by 4 h reoxygenation,respectively.The survival rate of cells was detected by methyl thiazol tetrazolium (MTT) chromatometry and the apoptosis was analyzed by flow cytometry.Results The cell survival rate in control group was (97.3 ±6.8)％,and of H/R group,which decreased significantly,was only (71.4 ± 5.9)％;there was significant difference between the UW group and control group (P ＜0.01);The cell survival rate in ARA290 group [(89.5 ± 7.1) ％] was slightly lower than that in control group (P ＜ 0.05),but significantly higher than in UW group and H/R group (P ＜ 0.01).The cell survival rate in UW group was (76.3 ± 6.0) ％,but there was no significant difference between the UW group and H/R group (P ＞ 0.05).The apoptosis rate in H/R group,control group,ARA290 group and UW group was respectively (13.13 ± 0.94) ％,(1.15 ± 0.18) ％,(5.34 ± 0.61) ％ and (11.45 ± 0.71) ％.There was statistically significant difference between ARA290 pretreatment group and the rest three groups (P ＜0.05).Conclusion Erythropoietin analogue ARA290 can show protective effect in HUVEC induced by H/R injury under hypothermic condition.And it can inhibit the apoptosis.