全氟化碳对脂多糖诱导肺微血管内皮细胞表达ICAM-1的影响

Effects of perfluorocarbon on lipopolysaccharide-induced intercellular adhesion molecule-1 expression in cultured pulmonary microvascular endothelial cells

目的观察全氟化碳(PFC)对脂多糖(LPS)诱导肺微血管内皮细胞(PMVEC)表达细胞间黏附分子-1(ICAM-1)的影响,探讨PFC的抗炎作用及其对PMVEC的保护机制。方法采用组织块法培养大鼠PMVEC,将细胞接种于Transwell小室,按随机数字表法随机分为空白对照组(C)、PFC干预组(F)、LPS刺激组(L)、LPS刺激+PFC干预组(LF),以处理0 h为空白对照,各处理组分别给予PFC或100 ng/ml LPS共孵育3、8、24 h,通过MTT法观测PFC作用后正常大鼠PMVEC细胞活力变化,应用逆转录PCR(RT-PCR)和流式细胞术(FCM)分别检测各组各时点大鼠PMVECICAM-1 mRNA和蛋白表达量。结果PFC作用后,正常大鼠PMVEC的细胞活力未见明显改变。各组各时点ICAM-1 mRNA表达的变化规律与蛋白质水平基本一致,F组各时点ICAM-1的表达量与C组比较均无显著性差异(P>0.05);L组各时点细胞的ICAM-1表达均较C组显著增强(P<0.05,P<0.01);LF组与L组同时点比较,3 h组ICAM-1表达量无明显差异(P>0.05),但8 h和24 h组均有显著降低(P<0.01,P<0.05)。结论PFC通过抗炎效应直接下调LPS诱导PMVEC表达ICAM-1,发挥PMVEC保护作用。

Objective To investigate the effects of perfluorocarbon（PFC） on lipopolysaccharide（LPS）-induced intercellular adhesion molecule-1（ICAM-1） expression in cultured pulmonary microvascular endothelial cells（PMVECs）.Methods Rat PMVECs cultured by peripheral lung tissue-sticking method were seeded in Transwell chambers and divided randomly into 4 groups： group C,remaining untreated as blank control;group F,incubated with PFC;group L,stimulated with 100 ng/ml LPS;group LF,treated with 100 ng/ml LPS and PFC.After treatment for 3,8 and 24 h,the cells in each group were harvested respectively.The cell viability of normal rat PMVECs treated by PFC was determined by MTT assay.Then the mRNA expression of ICAM-1 was semi-quantified by reverse transcription-polymerase chain reaction and the level of ICAM-1 protein expression was detected by flow cytometry.Results The cell viability of normal rat PMVECs was not impaired significantly by treatment of PFC.No signifant difference in mRNA and protein expressions of ICAM-1 was found between group F and group C（P0.05）.Treatment of cells in group L with 100 ng/ml LPS significantly induced the mRNA and protein expressions of ICAM-1 in a time-dependent manner（P0.05,P0.01）.In comparison with those in group L correspondingly,the mRNA and protein expressions of ICAM-1 in group LF were not inhibited significantly at 3 h after treatment（P0.05）,but were inhibited at 8 and 24 h after treatment（P0.01,P0.05）.Conclusion Perfluorocarbon suppresses the mRNA and protein expressions of ICAM-1 induced by LPS in cultured PMVECs,suggesting a potential mechanism that PFC protects PMVECs from LPS-induced injury via its anti-inflammatory effect.