国际麻醉学与复苏杂志
國際痳醉學與複囌雜誌
국제마취학여복소잡지
International Journal of Anesthesiology and Resuscitation
2015年
9期
805-809
,共5页
傅婕%江泽宇%封光%刘功俭
傅婕%江澤宇%封光%劉功儉
부첩%강택우%봉광%류공검
EphA2%抑制剂%内毒素%血管内皮细胞%凋亡
EphA2%抑製劑%內毒素%血管內皮細胞%凋亡
EphA2%억제제%내독소%혈관내피세포%조망
EphA2%Inhibiter%Lipopolysaccharides%Microvascular endothelial cells%Apoptposis
目的 探讨EphA2在内毒素(lipopolysaccharides,LPS)所致人肺血管内皮细胞(human pulmonary microvascular endothelial cells,HPMVECs)凋亡模型中的表达与作用. 方法 体外培养HPMVECs(24瓶),采用完全随机分组法分为4组(每组6瓶):对照组(C组)、EphA2抑制剂组(Fc组)、LPS干预组(LPS组)和LPS+EphA2抑制剂组(LPS+Fc组).流式细胞仪Annexin V/PI法检测每组细胞凋亡率,酶联免疫吸附法(enzyme-linked immunosorbent assay,ELISA)检测各组细胞上清液中肿瘤坏死因子(tumor necrosis factor,TNF)-α水平,Western blot检测各组EphA2水平. 结果 与C组比较,LPS组细胞的凋亡率升高[(7.8±1.4)％,(23.1±2.9)％](P＜0.05),TNF-α的量显著增加[(12.4±2.6),(37.1 ±5.3) ng/L] (P＜0.05),EphA2蛋白表达水平明显升高[(1.118±0.093),(2.244±0.127)](P＜0.05);与LPS组比较,LPS+Fc组细胞凋亡率下降[(23.1±2.9)％,(15.1±1.5)％](P＜0.05),TNF-α的量显著降低[(37.1±5.3),(25.5±2.9) ng/L](P＜0.05),EphA2表达明显下降[(2.244±0.127),(1.502±0.063)](P＜0.05);而C组与Fc组比较,细胞凋亡率[(7.8±1.4)％,(7.4±1.3)％]、TNF-α的量[(12.4±2.6),(14.2±2.3) ng/L]、EphA2表达量[(1.118±0.093),0.943±0.014)]差异无统计学意义(P＞0.05). 结论 LPS诱导的HPMVECs炎性反应凋亡模型中,抑制EphA2的表达可以减轻细胞凋亡.
目的 探討EphA2在內毒素(lipopolysaccharides,LPS)所緻人肺血管內皮細胞(human pulmonary microvascular endothelial cells,HPMVECs)凋亡模型中的錶達與作用. 方法 體外培養HPMVECs(24瓶),採用完全隨機分組法分為4組(每組6瓶):對照組(C組)、EphA2抑製劑組(Fc組)、LPS榦預組(LPS組)和LPS+EphA2抑製劑組(LPS+Fc組).流式細胞儀Annexin V/PI法檢測每組細胞凋亡率,酶聯免疫吸附法(enzyme-linked immunosorbent assay,ELISA)檢測各組細胞上清液中腫瘤壞死因子(tumor necrosis factor,TNF)-α水平,Western blot檢測各組EphA2水平. 結果 與C組比較,LPS組細胞的凋亡率升高[(7.8±1.4)％,(23.1±2.9)％](P＜0.05),TNF-α的量顯著增加[(12.4±2.6),(37.1 ±5.3) ng/L] (P＜0.05),EphA2蛋白錶達水平明顯升高[(1.118±0.093),(2.244±0.127)](P＜0.05);與LPS組比較,LPS+Fc組細胞凋亡率下降[(23.1±2.9)％,(15.1±1.5)％](P＜0.05),TNF-α的量顯著降低[(37.1±5.3),(25.5±2.9) ng/L](P＜0.05),EphA2錶達明顯下降[(2.244±0.127),(1.502±0.063)](P＜0.05);而C組與Fc組比較,細胞凋亡率[(7.8±1.4)％,(7.4±1.3)％]、TNF-α的量[(12.4±2.6),(14.2±2.3) ng/L]、EphA2錶達量[(1.118±0.093),0.943±0.014)]差異無統計學意義(P＞0.05). 結論 LPS誘導的HPMVECs炎性反應凋亡模型中,抑製EphA2的錶達可以減輕細胞凋亡.
목적 탐토EphA2재내독소(lipopolysaccharides,LPS)소치인폐혈관내피세포(human pulmonary microvascular endothelial cells,HPMVECs)조망모형중적표체여작용. 방법 체외배양HPMVECs(24병),채용완전수궤분조법분위4조(매조6병):대조조(C조)、EphA2억제제조(Fc조)、LPS간예조(LPS조)화LPS+EphA2억제제조(LPS+Fc조).류식세포의Annexin V/PI법검측매조세포조망솔,매련면역흡부법(enzyme-linked immunosorbent assay,ELISA)검측각조세포상청액중종류배사인자(tumor necrosis factor,TNF)-α수평,Western blot검측각조EphA2수평. 결과 여C조비교,LPS조세포적조망솔승고[(7.8±1.4)％,(23.1±2.9)％](P＜0.05),TNF-α적량현저증가[(12.4±2.6),(37.1 ±5.3) ng/L] (P＜0.05),EphA2단백표체수평명현승고[(1.118±0.093),(2.244±0.127)](P＜0.05);여LPS조비교,LPS+Fc조세포조망솔하강[(23.1±2.9)％,(15.1±1.5)％](P＜0.05),TNF-α적량현저강저[(37.1±5.3),(25.5±2.9) ng/L](P＜0.05),EphA2표체명현하강[(2.244±0.127),(1.502±0.063)](P＜0.05);이C조여Fc조비교,세포조망솔[(7.8±1.4)％,(7.4±1.3)％]、TNF-α적량[(12.4±2.6),(14.2±2.3) ng/L]、EphA2표체량[(1.118±0.093),0.943±0.014)]차이무통계학의의(P＞0.05). 결론 LPS유도적HPMVECs염성반응조망모형중,억제EphA2적표체가이감경세포조망.
Objective To investigate the expression and the role of EphA2 in lipopolysaccharides (LPS) induced apoptosis model of human pulmonary microvascular endothelial cells (HPMVECs).Methods HPMVECs were cultivated in vitro,and were randomly divided into four groups:control group (group C),Fc group,LPS group,LPS+Fc group.Apoptosis was detected by flow cytometry Annexin-FITC and PI double staining,western blot was performed to detect protein EphA2 expression in human pulmonary microvascular endothelial cells,and the concentration of tumor necrosis factor (TNF)-α was determined by enzyme-linked immunosorbent assay(ELISA).Results Compared with group C,apoptosis rate was significantly increased[(7.8±1.4)％,(23.1±2.9)％] in LPS group (P＜0.05).TNF-α concentration in LPS group was increased compared with group C [(12.4±2.6),(37.1±5.3) ng/L](P＜0.05).EphA2 protein expression of LPS group was increased [(1.118±0.093),(2.244±0.127)] compared with group C (P＜0.05).Compared with LPS group,apoptosis rate was significantly decreased[(23.1 ±2.9)％,(15.1±1.5)％] in LPS+Fc group(P＜0.05),TNF-α concentration was significantly decreased in LPS+Fc group [(37.1±5.3),(25.5±2.9) ng/L] (P＜0.05),and EphA2 protein expression in LPS+Fc group was significantly decreased [(2.244±0.127),(1.502±0.063)] (P＜0.05).Compared with group C,no statistically difference was found in apoptosis rate[(7.8±1.4)％,(7.4±1.3)％](P＞0.05),in TNF-α concertration[(12.4±2.6),(14.2±2.3) ng/L] (P＞0.05),in EphA2 protein expression [(1.118±0.093),(0.943±0.014)](P＞0.05)of Fc group.Conclusions In LPS-induced HPMVECs lung inflammation and apoptosis models,inhibiting the expression of EphA2 can reduce LPS-induced cell apoptposis.
