丙泊酚对体外内毒素激活人中性粒细胞NF-κB亚单位p65蛋白的影响

Effects of propofol on in vitro LPS-activated human polymorphonuclear nuclear transcription factor-kappaB subunit p65 protein

目的观察丙泊酚对内毒素(LPS)激活后人体中性粒细胞(PMN)NF-κB p65的表达及对PMN凋亡的影响。方法采集20例健康志愿者外周静脉血,分离PMN后分为五组:C组,加入生理盐水作为对照;LPS组,加入LPS 100ng/L);P1组,LPS+丙泊酚2μg/ml;P2组,LPS+丙泊酚4μg/ml;P3组,LPS+丙泊酚8μg/ml。孵育2.5h后,提取RNA;用RT-PCR法检测p65mRNA,Western blot检测p65蛋白含量,流式细胞术检测PMN凋亡。结果 LPS 100ng/L可以显著激活PMN,使p65表达大量增加。实验剂量的丙泊酚均可不同程度抑制LPS激活后PMN p65的表达(P<0.05);2-8μg/ml浓度的丙泊酚可剂量依赖性增加LPS导致的PMN凋亡(P<0.05或P<0.01)。结论丙泊酚2-8μg/ml可抑制PMN p65的激活,促进LPS激活后的PMN凋亡。

Objeetive To study the effects of propofol on human polymorphonuclear（PMN） cell nuclear transcription factor-κB （NF-κB） p65 expression and neutrophil apoptosis. Methods Peripheral blood samples of 20 healthy volunteers were collected and PMN cells were isolated, after which the PMN cells were divided into five groups and treated with normal saline （group C）, LPS 100 ng/L （group LPS）, LPS＋ propofol 2 μg/ml （ group P1 ）, LPS ＋ propofol 4 μg/ml （ group P2）, or LPS ＋ propofol 8 μg/ml（group P3）, respectively. After incubated for 2.5 hours, p65 mRNA expression was detected by RT-PCR, the expression of p65 protein was detected by Western blotting and PMN cell apoptosis was detected by flow cytometery. Results LPS 100 ng/L activated the expression of PMN NF-κB p65. Propofol 2-8μg/ml inhibited LPS-activated PMN NF-κB p65 expression（P〈0. 05） and promoted the apoptosis of LPS-induced PMN cells in a dose-dependent manner （P〈0. 05 or P〈0. 01）. Conclusion Propofol 2-8μg/ml inhibits LPS-activated PMN NF-κB p65 expression and promotes PMN cell apoptosis.