Relationship between the activation of heat shock factor and the suppression of nuclear factor-kappaB activity in rat hepatocyte cultures treated with cyclosporine A.

We investigated on primary cultures of rat hepatocytes the effect of cyclosporine A (CsA) on the activation of nuclear factor-kappaB (NF-kappaB), activator protein 1 (AP-1), and heat shock factor 1 (HSF1), three transcription factors involved in cellular response pathways. Hepatocytes were subjected to a time-course (1, 3, 6, and 22 hr) incubation and CsA treatment in the range 1-50 microM. NF-kappaB, AP-1, and HSF1 binding activities were established through electrophoretic mobility shift assay. Levels of HSP70 mRNA and protein were measured by Northern and Western blot analysis respectively. In cells incubated for 1 and 3 hr, electrophoretic mobility shift assay experiments showed a dose-dependent increase of the NF-kappaB binding activity; while following 22hr of incubation, a suppression of the positive effect of CsA at shorter times was detected. At all periods of incubation assayed, CsA induced the activation of AP-1 which was detected by DNA-binding activity of this transcription factor. A dose-dependent activation of HSF1 was observed at 22 hr of incubation. We conclude that in rat hepatocyte cultures, CsA induces the transcriptional activation of NF-kappaB, AP-1, and HSF1. However, the time point at which activation of each transcription factor occurs is different. Thus, at 22 hr of incubation, the CsA-induced activation of HSF1 is accompanied by the reduction of the positive effect of CsA on NF-kappaB activation at earlier time points.