摘要目的 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-BR mRNA表达的影响.方法 培养人脐静脉内皮细胞株ECV-304,以LPS 0.01、0.1、1、10 mg/L孵育ECV-304 2 h或用1 mg/L LPS孵育ECV-304 0.5、2、6、12 h,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1 mg/L LPS孵育细胞2 h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0.01 mg/L LPS诱导CCK-AR mRNA效应不明显,但可明显诱导CCK-BR mRNA表达;与1 mg/L LPS组比较,10 mg/L LPS孵育细胞2 h CCK-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,1 mg/L LPS孵育细胞0.5～2 h CCK-AR及CCK-BR的mRNA表达呈时间依赖性升高(P均<0.05);与LPS孵育2 h比较,1 mg/L LPS孵育细胞6 h CCK-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6 h比较,1 mg/L LPS孵育细胞12 h CCK-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 CCK-AR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.

abstractsObjective To investigate the expression of cholecystokinin-oetopeptide 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 rnRNA 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 rnRNA and CCK-BR mRNA.