高浓度二氧化碳预处理对人脐静脉内皮细胞缺氧复氧损伤的影响

目的 探讨高浓度二氧化碳(CO2)预处理对人脐静脉内皮细胞缺氧复氧损伤的影响.方法 人脐静脉内皮细胞接种于细胞培养板,随机分为6组,每组16孔,对照组(C组)常规培养;缺氧复氧组(MR组)缺氧4 h、复氧24 h;缺氧预处理组(APC组)缺氧10 min、复氧10 min,重复2次后缺氧4 h、复氧24 h;HCA.组、HCA2组和HCA3组分别于50%N-20%02-30%CO2培养箱中行30%CO2预处理10、30、60 min后常规培养10、20、30 min,缺氧4 h、复氧24 h.于复氧24 h时采用MTY法测定人脐静脉内皮细胞活力,采用免疫细胞化学法测定细胞间粘附分子-1(ICAM-1)表达水平.结果 与C组比较,MR组、APC组、HCA1～3组人脐静脉内皮细胞活力降低,ICAM-1表达上调(P＜0.05);与A/R组比较,APC组和HCA2组人脐静脉内皮细胞活力升高,APC组及HCA1,2组ICAM-1表达下调(P＜0.05).结论 高浓度CO2预处理可减轻人脐静脉内皮细胞缺氧复氧损伤.

Effects of hypercapnia preconditioning on anoxia/reoxygenation-induced injury to endothelial cells of human umbilical vein

Objective To investigate the effects of hypercapnia preconditioning on anoxia/reoxygenation (A/R)-induced injury to endothelial cells of human umbilical vein.Methods Cuhured endothelial cells of human umbilical vein purchased from the cell bank of Central South University Xiangya Medical College were incubated in 16 well culture plates and randomly divided into 6 groups(n=16 each):group Ⅰ control(C);groupⅡA/R;group Ⅲ anoxia preconditioning(APC);groupⅣ,Ⅴ,Ⅵ 10,30 and 60 min 30%CO2 preconditioning (HCA1,2,3).In group MR the endothehal cells were exposed to 95%N2-5%CO2 in an air-tight incubator at 37℃for 4 h followed by 24 h reoxygenation(75%N2-20%02-5%CO2 at 37℃).In group APC the endothehal cells were exposed to 10 min anoxia twice at 10 rain interval before MR.In group HCA1,2,3 the endothelial cells were exposed to 30%CO2 for 10,30 and 60 min respectively before A/R.At the end of 24 h reoxygenation the cell vitality(by MTT)and the ICAM-1 expression(by immuno-histochemistry)were meagnred.Results The cell vitality of the endothelial ceils was significantly decreased while ICAM-1 expression was up-regulated in group A/R (Ⅱ),APC(Ⅲ)and HCA1-3(Ⅳ,Ⅴ,Ⅵ)compared with group C(P＜0.05).The cell vitality of endothelial cells was significandy increased in group APC(Ⅲ)and group HCA2(Ⅴ)and ICAM-1 expression was downregulated in group APC(Ⅲ)and group HCA1,2(Ⅳ,Ⅴ)compared with group A/R(Ⅱ).Conclusion Hypercapnia preconditioning can ameliorate the MR injury to endothelial cells of human umbilical vein.