环氧化酶-2抑制剂对体外缺氧大鼠心肌细胞凋亡的影响  

Experimental study of effects of cyclooxygenase-2 inhibitor on apoptosis of hypoxic myocardial cells in vitro

作　　者：尹明[1] 沈洪[2] 黎檀实[2] 李银平[3] 刘刚[2] 

机构地区：[1]解放军总医院老年心血管病研究所,北京100853 [2]解放军总医院急诊科,北京100853 [3]天津市天和医院,300050

出　　处：《中国危重病急救医学》2006年第4期229-232,共4页Chinese Critical Care Medicine

基　　金：全军"十五"科研基金面上项目(01MA098)

摘　　要：目的 观察体外培养心肌急性缺氧条件下环氧化酶-2（COX2）抑制剂与细胞凋亡间的关系.方法 分离、培养Wistar大鼠心肌细胞.以第4～6代心肌细胞24份样本（每份1×10^5个细胞）随机分为正常对照组、单纯缺氧组、缺氧＋COX-2抑制剂（NS-398,20 μmol/L）组及缺氧＋阿司匹林（100 μg/L）组.缺氧前30 min加入干预药物或等量二甲亚砜.以蛋白质免疫印迹法检测心肌细胞中COX-1/2表达;流式细胞仪检测心肌细胞凋亡率;放射免疫法（RIA）检测心肌细胞培养液中6-酮-前列腺素F1α（6-ketoPGF1α）、血栓素B2（TXB2）含量.结果 各组心肌细胞中COX-1表达差异无显著性;COX2表达均较正常对照组增高,阿司匹林与NS-398均不抑制COX2表达.心肌细胞凋亡率由高到低依次为缺氧＋NS-398组、缺氧＋阿司匹林组、单纯缺氧组和正常对照组;缺氧＋NS-398组与其他各组间比较差异均有显著性（P均〈0.05）.各组心肌细胞培养液内TXB2水平由低到高依次为正常对照组、缺氧＋NS-398组、缺氧＋阿司匹林组和单纯缺氧组;缺氧＋NS-398组与单纯缺氧组比较差异有显著性（P〈0.05）;6-keto-PGF1α水平由低到高依次为正常对照组、缺氧＋NS-398组、缺氧＋阿司匹林组和单纯缺氧组;缺氧＋NS-398组与单纯缺氧组比较差异有显著性（P〈0.05）;TXB2/6-keto-PGF1α比值差异无显著性.结论 急性缺氧可直接诱导培养心肌细胞表达COX-2,而COX-1表达不增加,缺氧使培养液内COX-2作用产物TXB2、6keto-PGF1α水平升高,此效应无中性粒细胞、巨噬细胞等炎症细胞参与.COX-2抑制剂NS-398可降低缺氧心肌培养液内TXB2、6-keto-PGF1α升高程度,但不改变TXB2/6keto-PGF1α比值.急性缺氧可直接诱导心肌细胞凋亡,NS398可显著增加缺氧培养心肌凋亡率,此效应不需要其他组织细胞参与.Objective To observe the effects of cyclooxygenase （COX）- 2 inhibitor on apoptosis of hypoxic myocardial cells in vitro. Methods The myocardial cells were obtained from new-born Wistar rats, and were dispersed to single cell with trypsin. The cells in 4th - 6th passages were randomly divided into 24 samples （every sample contained 1×10^5 cells）： normal control group, hypoxic myocardial cells control group ; hypoxic myocardial cells ＋ NS - 398 （20μmol/L ） and hypoxic myocardial cells ＋ aspirin （100μg/L）. During culture, oxygen was replaced by N2, and the cells were cultivated in 5% CO2＋95% N2 at 37℃ for 6 hours. Either interventional medicine or same amount of dimethyl sulphoxide （menstruum） was added to the cells 30 minutes before hypoxia. The expression of COX - 1 and COX - 2 in cultured myocardial cells in vitro was examined with Western blot. The apoptosis percentage of cells in each group was examined with flow cytometry. After centrifuging the culture medium under low temperature, 6 -keto-prostaglandin F1α. （6- keto- PGF1α,） and thromboxane B2 （TXB2） were determined with radioimmunoassay （RIA）. Results There was no significant difference in the expression of COX- 1 among the groups. The expression of COX - 2 was higher in all acute hypoxic myocardial cells groups compared with the control group. Neither aspirin nor NS - 392 inhibited the expression of COX - 2. The positive percentage of apoptosis of cultured myocardial ceils ranked as follow： hypoxic＋NS- 398 group, hypoxic ＋aspirin group, hypoxic control group and normal control group. The differences of apoptosis rate between hypoxic ＋ NS - 398 group and other groups were significant （all P〈0. 05）. The levels of TXB2 in each group of cell medium ranked in the following order： normal control group, hypoxic ＋NS - 398 group, hypoxic ＋aspirin group and hypoxic control group. The difference of TXB2 level between hypoxic＋NS - 398 group and hypoxic control group was significant （P〈0.05）. Th

关 键 词：环氧化酶-2抑制剂 心肌 缺氧 细胞凋亡 

分 类 号：R965[医药卫生—药理学]