Title: Effects of thiopental pretreatment on NF-κB activity in TNF-α activated Jurkat cells
Abstract: Objective To investigate the effects of thiopental (TP) pretreatment on nuclear factor-κB (NF-κB) activity in TNF-ακ activated Jurkat cells. Methods Jurkat cells of a human T lymphoma cell line (from incubate preservation center, Wuhan University) were used. The Jurkat cells were incubated in 5% CO2 incubator at a concentration of 1×105·ml-1 at 37℃ for 24h. The experiment was performed in 2 parts. In part A the cells were divided into 6 groups: group A1 blank control; group A2 TNF-α S U·ml-1; group A3 TP400 μg·ml-1; group A4 TP 1 000 μg·ml-1; group A5 TP 400 μg·ml-1 + TNF-α 5 U·ml-1 and group A6 TP 1 000 μg·ml-1 + TNF-α 5 U·ml+1 . In group AS and A6 the cells were incubated first in the presence of TP 400 or 1 000 μg·ml-1 for 1 h. The cells were then washed and incubated in the medium containing TNF-α 5 U·ml-1 for 1h. The nuclear protein of the cells was then collected for determination of NF-κB activity using electrophoretic mobility shift assay (EMSA) and expression IκB-α protein (an inhibitory protein of NF-κB) using Western blotting. In part B the cells were divided into 8 groups: group B1 blank control; group B2 TNF-α 5 U·ml-1; group B 3-8 in which the cells were incubated in the presence of TP 1 000 μg·ml-1 for 1 h and then washed and incubated in the presence of TNF-α S U·ml-1 for 1 h immediately (B3) or after 2, 4, 6, 7, 8 h (B4-8). The cells were then collected for determination of NF-κB activity. Results The NF-κB activity was significantly increased and LκB-α expression significantly down-regulated in the TNF-α activated Jurkat cells as compared with the blank control. The increase in NF-κB activity and down-regulation of IκB-α protein expression in the Jurkat cells induced by TNF-α could be inhibited by pretreatment with thiopental 1 000 μg·ml-1. The inhibitory effect of thiopental pretreatment persisted up to 8 hours after being washed off. Conclusion Thiopental pretreatment can inhibit the increase in NF-κB activity in Jurkat cells induced by TNF-α. The inhibitory effect of thiopental pretreatment may be related to up-regulation of IκB-α expression and persists up to 8 hours.
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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