Effect of travoprost on nuclear factor kappa B expression in human ciliary muscle cells

Jianhui XIAO; Yuqing LAN; Chi ZHANG; Zhaoxia XIA; Wei PENG

ABSTRACT

BACKGROUND:

Nuclear factor kappa B (NF-κB) is possibly related to regulation of various cell signals that are derived from aqueous uveoscleral outflow pathway.

OBJECTIVE:

To explore effect of travoprost on the expression of NF-κB and inhibitor-κB (I-κB) in human ciliary muscle cells cultured in vitro. DESIGN, TIME AND

SETTING:

A contrast study, which was performed in the Laboratory of Zhongshan Ophthalmology Center from March 2005 to November 2006.MATERIALS Eyeballs were obtained from the youth who died due to other diseases except eye disease no more than one hour. The relatives voluntarily provided the informed consent.

METHODS:

Travoprost (1 μmol/L) was added in human ciliary muscle cell culture medium, and then the samples were divided into four groups according to culture time, including 0-hour (control group), 6-hour, 12-hour, and 24-hour experimental groups. MAIN OUTCOME

MEASURES:

Expression of mRNA and protein of NF-κB p65 and I-κBα in the four groups by using real-time RT-PCR, immunofluorescence relative quantitative analysis and enzyme linked immunosorbent assay (ELISA) techniques.

RESULTS:

As compared to control group, mRNA expression of NF-κB p65 in the 6-hour, 12-hour, and 24-hour experimental groups was decreased (F=17.068, P=0.001); while mRNA expression of I-κBα was not changed remarkably in the 6-hour and 12-hour experimental groups (P > 0.05), but the expression was significantly higher than that in the 24-hour experimental group (F=32.742, P=0.000). Immunofluorescence relative quantitative analysis showed that the fluorescence intensity of NF-κB p65 in the 6-hour, 12-hour, and 24-hour experimental groups were weaker than that in the 0-hour control group (F=17.216, P=0.000); additionally, as compared to 0-hour control group, fluorescence intensity of I-κBα in the 6-hour experimental group was not changed remarkably (P=0.134), that in the 12-hour experimental group was weakened (P=0.032), and that in the 24-hour experimental group was strengthened (F=17.346, P=0.001). ELISA revealed that expression of phosphorylated NF-κB p65 was decreased gradually by the time of being induced by travoprost (F=15.4, P=0.001).

CONCLUSION:

Travoprost can down-regulate mRNA expression of NF-κB p65, inhibit nuclear translocation, and up-regulate mRNA expression of I-κBα in human ciliary muscle cells.