CAJ | 학술논문

目的：观察阿折地平对西罗莫司洗脱支架致血管内皮损伤的干预作用，并探讨其作用机制。方法培养人脐静脉内皮细胞（ HUVEC），将HUVEC分为对照组、模型组和实验组。对照组不加药物，模型组给予西罗莫司500 nmol／L，实验组给予阿折地平10μmol／L＋西罗莫司500 nmol／L。各组处理24 h后，HE染色观察细胞形态变化，测定培养液中的NO，Fluo-3／AM荧光探针标记细胞内Ca2＋，JC-1荧光探针检测线粒体膜电位，Annexin V FITC／PI染色法测算细胞凋亡率。结果各组处理24 h后，对照组细胞呈多角形、单层铺路石样排列；模型组细胞肿胀，胞质出现空泡，部分细胞核固缩、碎裂；实验组细胞基本呈多角形、单层铺路石样排列，少见细胞肿胀及胞质空泡。与对照组相比，模型组与实验组细胞培养液中NO水平降低、胞质Ca2＋浓度升高、细胞凋亡率升高、线粒体膜电位下降（P均＜0．05）；与模型组相比，实验组细胞培养液中NO水平升高、胞质Ca2＋浓度及细胞凋亡率降低、线粒体膜电位升高（P均＜0．05）。结论阿折地平可在一定程度上抑制西罗莫司洗脱支架导致的血管内皮损伤，其机制可能与提高细胞内NO水平、降低细胞内Ca2＋浓度、抑制线粒体膜电位降低、减少细胞凋亡有关。
목적：관찰아절지평대서라막사세탈지가치혈관내피손상적간예작용，병탐토기작용궤제。방법배양인제정맥내피세포（ HUVEC），장HUVEC분위대조조、모형조화실험조。대조조불가약물，모형조급여서라막사500 nmol／L，실험조급여아절지평10μmol／L＋서라막사500 nmol／L。각조처리24 h후，HE염색관찰세포형태변화，측정배양액중적NO，Fluo-3／AM형광탐침표기세포내Ca2＋，JC-1형광탐침검측선립체막전위，Annexin V FITC／PI염색법측산세포조망솔。결과각조처리24 h후，대조조세포정다각형、단층포로석양배렬；모형조세포종창，포질출현공포，부분세포핵고축、쇄렬；실험조세포기본정다각형、단층포로석양배렬，소견세포종창급포질공포。여대조조상비，모형조여실험조세포배양액중NO수평강저、포질Ca2＋농도승고、세포조망솔승고、선립체막전위하강（P균＜0．05）；여모형조상비，실험조세포배양액중NO수평승고、포질Ca2＋농도급세포조망솔강저、선립체막전위승고（P균＜0．05）。결론아절지평가재일정정도상억제서라막사세탈지가도치적혈관내피손상，기궤제가능여제고세포내NO수평、강저세포내Ca2＋농도、억제선립체막전위강저、감소세포조망유관。
Objective To investigate the protective effect of calcium-channel blockers azelnidipine on sirolimus elu-ting stent-induced vascular endothelial injury and its mechanism .Methods Human umbilical vein endothelial cells ( HU-VECs) were cultivated and divided into the control group (treated with culture medium), model group (treated with siroli-mus 500 nmol/L) and the experimental group (azelnidipine 10 μmol/L +sirolimus 500 nmol/L), respectively.After 24 h of treatment, changes in cell morphology were observed by HE staining .Effects on the production of nitric oxide (NO) were detected by Nitric Oxide Assay Kit .The intracellular calcium ion ( Ca2+) concentration was assayed with Fluo-3/AM staining, the changes of mitochondrial membrane potential was detected by JC -1 fluorescence labeling , and the apoptosis rate of HUVECs was analyzed by Annexin V FITC/PI staining.Results After 24 h of treatment, in the control group, cells were polygonal and in the single cobblestone arrangement;in the model group, cell swelled, cytoplasm had vacuoles, part of the nucleus had pycnosis , and nuclear fragmentation were observed;in the experimental group , cells were substan-tially polygonal , and in the single cobblestone arrangement , the cell swelling and cytoplasmic vacuoles were rare .Com-pared with the control group , the levels of NO in the cell culture fluid were reduced , the levels of intracellular free Ca 2+were increased , apoptosis rate was increased and mitochondrial membrane potential was reduced in the model group and ex -perimental group (all P<0.05).Compared with the model group, the levels of NO in the cell culture fluid were in-creased , the levels of intracellular free Ca 2+were reduced , the apoptosis rate was reduced and the mitochondrial membrane potential was increased in the experimental group (all P<0.05).Conclusions Azelnidipine has protective effect on sirolimus eluting stent-induced vascular endothelial injury .The possible mechanism might be related to the decrease of in-tracellular Ca 2+which could alleviate calcium overload and mitochondrial membrane potential and finally reduce apoptosis .