八肽缩胆囊素受体在血管内皮细胞中的表达及脂多糖的影响

The expression of cholecystokinin-octopeptide receptor in vascular endothelial cells induced by lipopolysaccharide

目的观察八肽缩胆囊素受体（CCK—R）mRNA在人脐静脉内皮细胞株ECV-304中的表达，并探讨内毒素脂多糖（LPS）对CCK—AR、CCK—BRmRNA表达的影响。方法培养人脐静脉内皮细胞株ECV-304，以LPS0．01、0．1、1、10mg／L孵育ECV-3042h或用1mg／LLPS孵育ECV-304 0．5、2、6、12h，采用逆转录-聚合酶链反应（RT—PCR）检测CCK—AR、CCK—BR的mRNA表达，并对扩增产物的特异性进行测序分析。结果与空白对照组比较，0．01、0．1及1mg／LLPS孵育细胞2h可剂量依赖性引起CCK-AR及CCK—BR的mRNA表达明显升高（P均〈0．05），其中0．01mg／LLPS诱导CCK—AR mRNA效应不明显，但可明显诱导CCK—BR mRNA表达；与1mg／LLPS组比较，10mg／LLPS孵育细胞2hCCK—AR及CCK—BR的mRNA表达未继续出现明显升高（P均〉0．05）。与空白对照组比较，1mg／LLPS孵育细胞0．5～2h CCK—AR及CCKBR的mRNA表达呈时间依赖性升高（P均〈0．05）；与LPS孵育2h比较，1mg／LLPS孵育细胞6hCCK—AR及CCK—BR的mRNA表达明显下降，但仍高于空白对照组（P均〈0．05）；与LPS孵育6h比较，1mg／LLPS孵育细胞12hCCK—AR及CCK—BR的mRNA表达进一步降低（P均〈0．05），且与空白对照组比较已无显著差异（P均〉0．05）。结论CCKAR与CCK—BR的mRNA在ECV-304中均有表达；LPS可诱导CCK—AR及CCK—BR的mRNA表达上调。

Objective To investigate the expression of cholecystokinin-octopeptide receptor （CCK-R） mRNA, and observe the effect of lipopolysaccharide （LPS） on CCK-AR mRNA and CCK-BR mRNA expression in ECV-304. Methods The human umbilical vein endothelial cell line ECV-304 was cultured and treated with LPS in dosage of 0.01, 0.1, 1, 10 mg/L for 2 hours, or treated with LPS in dosage of 1 mg/L for 0. 5, 2, 6, 12 hours. Reverse transcription-polymerase chain reaction （RT-PCR） was employed to examine the expression of CCK-AR mRNA and CCK-BR mRNA in ECV-304, and to analyse the sequences of the amplification products. Results Compared with control group, the expression of CCK-AR mRNA and CCK-BR mRNA was significantly upregulated in a dose-dependence manner after incubated with 0.01, 0.1 and 1 mg/L LPS for 2 hours （all P〈0.05）. However, the expression of CCK-AR mRNA showed no significant increase ,while that of CCK-BR mRNA was increased, after being incubated with 0.01 mg/L LPS. The expressions of CCK-AR mRNA and CCK-BR mRNA in the 10 mg/L LPS group showed no significant difference compared with 1 mg/L LPS group （both P 〉 0.05）. The expression of CCK-AR mRNA and CCK-BR mRNA was significantly upregulated in a time dependence manner after incubated with 1 mg/L LPS from 0.5 hour to 2 hours compared with control group （all P〈0.05）. After incubated with 1 mg/L LPS for 6 hours, the expression of CCK-AR mRNA and CCK-BR mRNA was significantly decreased compared with 2 hour group, but was still higher than that of control group （both P〈0.05）. Its expression was decreased further after being incubated with 1 mg/L LPS for 12 hours compared with the 6 hours group （both P〈0.05）, but showed no significant difference compared with the control group （both P〉0.05）. Conclusion Both CCK AR mRNA and CCK-BR mRNA are expressed in ECV-304. LPS can up-regulate the expression of CCK AR mRNA and CCK-BR mRNA.