摘要:Objective: To observe the effect of serum of sepsis patients on the expression of Nuclear Factor-Kappa B (NF-kB) on human pulmonary vascular endothelial cells ( PMVECs) , and to explore the mechanism of severe sepsis induced acute lung injury (ALI). Methods; PMVECs were cultured in vitro, and were divided into 3 groups randomly: normal culture group (group A) , healthy serum group (group B) , patients'serum group (group C). The expressions of NF-kB were obseverd and values of average optical density (AOD) were detected by immunohistochemistry at 0, lh, 2h, 4h, 6h after stimulation. Concentration of TNF-a was detected by ELISA. The above indexes of pulmonary vascular endothelial cells were compared between 3 groups. Results: Compared with that of group A and group B, the expression of NF-kB in PMVECs showed strongly positive in group C, and AOD values reached peak at 1 hour, and decreased gradually (0. 523 + 0.012, 0. 498 ±0.009, 0. 444 ±0. 030). No significant difference of TNF-α between group A and group B was found (P >0.05). Compared with that of group A and group B, the level of TNF-α was significantly increased at each time point, and showed significant differences (P<0.01). Conclusion:Serum of severe sepsis patients can activate NF-kB and thus initiate the secretion of TNF-α, which may be an important pathway of ALI caused by severe sepsis.%目的:观察严重脓毒症患者血清对人肺血管内皮细胞(pulmonary microvascular endothelial cell,PMVECs) NF-kB表达的影响,探讨严重脓毒症致急性肺损伤的发生机制.方法:随机将离体培养PMVECs分为3组:正常培养组(加入10%胎牛血清,A组)、健康血清组(加入10%健康人血清,B组)、患者血清组(加入10%严重脓毒症患者血清,C组),分别于刺激0、1、2、4、6h免疫组化观察离体PMVECs NF-κB表达并测定各组平均光密度(average optical density,AOD)值;ELISA法检测培养上清液分泌的TNF-α水平;比较不同时相3组PMVECs上述指标变化.结果:与A组及B组相比,C组PMVECs胞核NF-κB呈强阳性表达,且1 h AOD值达高峰,后逐渐下降(0.523±0.012,0.498±0.009,0.444±0.030);ELISA显示:不同时相A组与B组TNF-α水平均无明显差异(P>0.05),C组与A组、B组相比各时间点TNF-α水平均明显升高,且均有显著差异(P<0.01).结论:严重脓毒症患者血清可通过活化NF-κB启动分泌TNF-α,这可能是严重脓毒症致急性肺损伤的一个重要通路.