Tumor necrosis factor-alpha enhances mRNA expression and secretion of interleukin-6 in cultured human airway smooth muscle cells
Airway smooth muscle (ASM) is considered to be an end-target cell for the effects of mediators released during airway wall inflammation. Several reports suggest that activated ASM may be capable of generating various proinflammatory cytokines. We investigated the effects of tumor necrosis factor (TNF)-alpha, a potent proinflammatory cytokine, on cultured human ASM cells by examining the expression and release of the cytokine interleukin (IL)-6, cell proliferation, and the expression pattern of c-fos and c-jun, two nuclear proto-oncogenes constituting the activator protein-1 transcription factor. Growth-arrested cell monolayers were stimulated with human recombinant TNF-alpha in a concentration- and time-dependent manner. TNF-alpha stimulated the expression of IL-6 messenger RNA (mRNA), which was detected after 15 min, reaching a maximum at 1 h. IL-6 protein was readily detected in ASM cell-conditioned medium after 2 h of TNF-alpha stimulation. Protein levels increased in a time- and concentration-dependent manner. Release of IL-6 elicited by TNF-alpha was significantly inhibited by dexamethasone, cycloheximide, and nordihydroguaiaretic acid (NDGA). TNF-alpha did not alter DNA biosynthesis up to 48 h or cell numbers up to 120 h. Northern blot analysis of proto-oncogene expression revealed that c-fos and c-jun mRNA levels were elevated after 30 min of TNF-alpha incubation with maximum levels at 1 h and 45 min, respectively. Expression of c-fos mRNA was downregulated by NDGA. Four hours of TNF-alpha treatment resulted in translocation of c-jun immunofluorescence from the cytoplasm to the nucleus in human ASM cells. Our results suggest that despite the lack of a mitogenic response to TNF-alpha, upregulation of primary response genes in human ASM cells may account for the induction of proinflammatory cytokines, such as IL-6, in human airways.
|Keywords||Bronchi/cytology/*drug effects/metabolism, Cell Division/drug effects, Cell Nucleus/drug effects/metabolism, Cells, Cultured, Cycloheximide/pharmacology, Cytoplasm/drug effects/metabolism, Dexamethasone/pharmacology, Genes, fos/genetics, Genes, jun/genetics, Humans, Immunohistochemistry, Interleukin-6/genetics/*metabolism, Lipoxygenase Inhibitors/pharmacology, Muscle, Smooth/cytology/*drug effects/metabolism, Nordihydroguaiaretic Acid/pharmacology, Proto-Oncogene Proteins c-jun/genetics/metabolism, RNA, Messenger/genetics/metabolism, Research Support, Non-U.S. Gov't, Tumor Necrosis Factor-alpha/*pharmacology, Up-Regulation/*drug effects|
McKay, S., Hirst, S.J., Bertrand-de Haas, M., de Jongste, J.C., Hoogsteden, H.C., Saxena, P.R., & Sharma, H.S.. (2000). Tumor necrosis factor-alpha enhances mRNA expression and secretion of interleukin-6 in cultured human airway smooth muscle cells. American Journal of Respiratory Cell and Molecular Biology. Retrieved from http://hdl.handle.net/1765/9390