Objective: To investigate the expression of cholecystokinin-octapeptide 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.