LPS及TNFα对肺微血管内皮细胞内游离钙影响.docVIP

LPS及TNFα对肺微血管内皮细胞内游离钙影响 　 作者：肖贞良，全燕，李福祥，钱桂生 【摘要】 目的 探讨脂多糖(LPS)、肿瘤坏死因子α(TNFα)对大鼠肺微血管内皮细胞(RPMVEC)内游离钙浓度([Ca2+]i)的影响。方法 将细胞接种于盖玻片上，细胞单层汇合后进行实验。实验分组：LPS组(用10μg/ml LPS)，LPS+山莨菪碱组(用10μg/ml LPS+10μg/ml山莨菪碱)，TNFα组(用5000u/mlTNFα)，TNFα+山莨菪碱组(用5000u/ml TNFα+10μg/ml山莨菪碱)，正常对照组加等量稀释液。作用15、90min后按照试剂盒提供的方法，并参照文献测定[Ca2+]i。结果 与正常对照组比较，LPS组、LPS+山莨菪碱组15min和90min两个时相点[Ca2+]i均显著升高(Plt;0.01);LPS组与LPS+山莨菪碱组比较，15min和90min两个时相点[Ca2+]i无明显差异(Pgt;0.05)。同样，与正常对照组比较，TNFα组、TNFα+山莨菪碱组15min和90min两个时相点[Ca2+]i均显著升高(Plt;0.01);TNFα组与TNFα+山莨菪碱组比较，15min和90min两个时相点[Ca2+]i无明显差异(Pgt;0.05)。结论 LPS、TNFα作用可使RPMVEC [Ca2+]i显著升高，山莨菪碱对LPS、TNFα诱导的RPMVEC [Ca2+]i升高无显著抑制作用;LPS、TNFα引起的[Ca2+]i浓度升高与β-AR介导的信号转导通路无关;[Ca2+]i升高可能参与了G蛋白偶联受激酶的活性调节。 【关键词】 内皮细胞;游离钙;脂多糖;肿瘤坏死因子α 　　Abstract Objective To investigate the effects of lipopolysaccharide (LPS) and tumor necrosis factor α (TNF α) on intracellular free calcium ([Ca2+]i) of rat pulmonary microvascular endothelial cells (RPMVEC) and the relationship between [Ca2+]i. and beta adrenoceptor (βAR).Methods Equal amount of cells was incubated on glass films; experiment was carried out when the cell monolayer confluented; group division in the experiment was as follows: LSP group (10μg/ml of LPS added), LPS+anisodamine group (10μg/ml of LPS and 10μg/ml of anisodamine added), TNFα group (5000μ/ml of TNFα added), TNFα+anisodamine group (5000μ/ml of TNFα and 10μg/ml of anisodamine added) and normal control group (equal amount of diluent added), and each group had 4 samples. [Ca2+]i. In each group was measured at the time point of 15 and 90 minute with the method from reagent kit instruction and literature.Result [Ca2+]i in LPS group and LPS+anisodamine group at the time point of 15 and 90 minutes were all significantly higher than that in normal control group (Plt;0.01), but there existed no significant difference of [Ca2+]i between LPS group and LPS+anisodamine group at the same time points (Plt;0.01); [Ca2+]i in TNFα group and TNFα+anisodamine group at the time point of 15