LPS和TNFα对肺微血管内皮细胞内游离钙的影响.doc

LPS和TNFα对肺微血管内皮细胞内游离钙的影响
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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();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()。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();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()。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 haride (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 aniso