脂多糖对肺动脉内皮细胞Ⅰ型Na^+/H^+交换器表达的影响  

The changes of NHE-1 by lipopolysaccharide on pulmonary artery endothelial cells

作　　者：赵澎涛[1] 李志超[1] 刘毅[1] 李剑[1] 

机构地区：[1]第四军医大学基础部病理生理学教研室,陕西西安710032

出　　处：《心脏杂志》2003年第3期199-201,204,共4页Chinese Heart Journal

摘　　要：目的 :探讨脂多糖 ( LPS)对培养的牛肺动脉内皮细胞 ( PAEC) 型 Na+ /H+交换器 ( NHE-1)表达的影响及其与内皮细胞死亡的关系。方法 :体外培养牛 PAEC,MTT比色检测细胞存活率 ,并提取总 RNA,应用反转录聚合酶链式反应 ( RT-PCR)及 DNA电泳技术 ,以 NHE-1与β-actin电泳条带光密度值之比 ( e/c)作数据分析 ,以其比值反映 NHE-1m RNA的表达程度。应用 Fluo3 /Amy荧光探针标记 ,激光共聚焦观察细胞内钙离子的浓度。结果 :在L PS( 0 .0 1～ 1.0 μg/ml)作用 48h后 ,MTT比色吸光度值分别为 0 .3 5± 0 .0 7,0 .3 1± 0 .0 7,0 .2 6± 0 .0 4,低于对照组 0 .44± 0 .0 6( P<0 .0 1)。e/c分别为 0 .43± 0 .11,0 .83± 0 .14 ,1.2 2± 0 .19,高于对照组 0 .2 8± 0 .0 8( P<0 .0 1,n=4)。激光共聚焦结果显示 L PS引起细胞内钙离子增多 ,呈剂量依赖型。结论 :LPS可以引起肺动脉内皮细胞NHE-1表达增加 ,抑制细胞增殖 ,导致细胞死亡 ,钙离子可能参与 NHE-1的激活及表达。AIM To study the changes of sodium hydrogen exchanger isoform 1 (NHE 1) on bovine pulmonary artery endothelial cells with the function of lipopolysaccharide (LPS), and the relationship with endothelial cells death. METHODS: Endothelial Cells were separated from bovine pulmonary artery and cultured in vitro. Cell activities were reflected by the OD value of MTT. Total RNA extraction and RT PCR method were used to measure the mRNA expression of NHE 1. Intracellular free Ca 2+ concentration was measured by calcium fluorescent probe Fluo3/AM and laser confocal microscope. RESULTS: In the presence of LPS(0.01～1.0 μg/ml), after 48 hours,the absorbencies of MTT(0.35±0.07,0.31±0.07,0.26±0.04) were lower in the experimental group than in the control group (0.44±0.06)( P <0.01),but e/c(0.43±0.11,0.83±0.14,1.22±0.19) were higher than control group (0.28±0.08) ( P <0.01, n =4). Fluoresent intensity of intracellular free Ca 2+ increased in a dose dependant manner after the addition of LPS. CONCLUSION: LPS can cause a increased expression of NHE 1 mRNA,which may play an important role in endothelial cells death and intracellular free Ca 2+ may participate in the activition and expression of NHE 1.

关 键 词：脂多糖 Ⅰ型Na^+/H^+交换器 肺动脉内皮细胞 钙离子 

分 类 号：R322.12[医药卫生—人体解剖和组织胚胎学]